Resistant Recombinant Inbred Lines Research Articles

Comparative Transcriptomic Analysis of Soybean Cyst Nematode Inbred Populations Non-adapted or Adapted on Soybean rhg1-a/Rhg4-Mediated Resistance.

Soybean cyst nematode (SCN, Heterodera glycines) is most effectively managed through planting resistant soybean cultivars, but the repeated use of the same resistance sources has led to a widespread emergence of virulent SCN populations that can overcome soybean resistance. Resistance to SCN HG type 0 (Race 3) in soybean cultivar Forrest is mediated by an epistatic interaction between the soybean resistance genes rhg1-a and Rhg4. We previously developed two SCN inbred populations by mass-selecting SCN HG type 0 (Race 3) on susceptible and resistant recombinant inbred lines, derived from a cross between Forrest and the SCN-susceptible cultivar Essex, which differ for Rhg4. To identify SCN genes potentially involved in overcoming rhg1-a/Rhg4-mediated resistance, we conducted RNA sequencing on early parasitic juveniles of these two SCN inbred populations infecting their respective hosts, only to discover a handful of differentially expressed genes (DEGs). However, in a comparison with early parasitic juveniles of an avirulent SCN inbred population infecting a resistant host, we discovered 59 and 171 DEGs uniquely up- or downregulated in virulent parasitic juveniles adapted on the resistant host. Interestingly, the proteins coded by these 59 DEGs included vitamin B-associated proteins (reduced folate carrier, biotin synthase, and thiamine transporter) and nematode effectors known to play roles in plant defense suppression, suggesting that virulent SCN may exert a heightened transcriptional response to cope with enhanced plant defenses and an altered nutritional status of a resistant soybean host. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Plant necrotrophic bacterial disease resistance phenotypes, QTL, and metabolites identified through integrated genetic mapping and metabolomics in Solanum species.

Most food crops are susceptible to necrotrophic bacteria that cause rotting and wilting diseases in fleshy organs and foods. All varieties of cultivated potato (Solanum tuberosum L.) are susceptible to diseases caused by Pectobacterium species, but resistance has been demonstrated in wild potato relatives including S. chacoense. Previous studies demonstrated that resistance is in part mediated by antivirulence activity of phytochemicals in stems and tubers. Little is known about the genetic basis of antivirulence traits, and the potential for inheritance and introgression into cultivated potato is unclear. Here, the metabolites and genetic loci associated with antivirulence traits in S. chacoense were elucidated by screening a sequenced S. tuberosum x S. chacoense recombinant inbred line (RIL) population for antivirulence traits of its metabolite extracts. Metabolite extracts from the RILs exhibited a quantitative distribution for two antivirulence traits that were positively correlated: quorum sensing inhibition and exo-protease inhibition, with some evidence of transgressive segregation, supporting the role of multiple loci and metabolites regulating these resistance-associated systems. Metabolomics was performed on the highly resistant and susceptible RILs that revealed 30 metabolites associated with resistance, including several alkaloids and terpenes. Specifically, several prenylated metabolites were more abundant in resistant RILs. We constructed a high-density linkage map with 795 SNPs mapped to 12 linkage groups, spanning a length of 1,507 cM and a density of 1 marker per 1.89 cM. Genetic mapping of the antivirulence and metabolite data identified five quantitative trait loci (QTLs) related to quorum sensing inhibition that explained 8-28% of the phenotypic variation and two QTLs for protease activity inhibition that explained 14-19% of the phenotypic variation. Several candidate genes including alkaloid, and secondary metabolite biosynthesis that are related to disease resistance were identified within these QTLs. Taken together, these data support that quorum sensing inhibition and exo-protease inhibition assays may serve as breeding targets to improve resistance to nectrotrophic bacterial pathogens in potato and other plants. The identified candidate genes and metabolites can be utilized in marker assisted selection and genomic selection to improve soft- rot and blackleg disease resistance.

Insights into the early transcriptomic response against watermelon mosaic virus in melon

Background Watermelon mosaic virus (WMV) is one of the most prevalent viruses affecting melon worldwide. Recessive resistance to WMV in melon has previously been reported in the African accession TGR-1551. Moreover, the genomic regions associated to the resistance have also been described. Nevertheless, the transcriptomic response that might infer the resistance to this potyvirus has not been explored.ResultsWe have performed a comparative transcriptomic analysis using mock and WMV-inoculated plants of the susceptible cultivar “Bola de oro” (BO) and a resistant RIL (Recombinant inbred line) derived from the initial cross between “TGR-1551” and BO. In total, 616 genes were identified as differentially expressed and the weighted gene co-expression network analysis (WGCNA) detected 19 gene clusters (GCs), of which 7 were differentially expressed for the genotype x treatment interaction term. SNPs with a predicted high impact on the protein function were detected within the coding regions of most of the detected DEGs. Moreover, 3 and 16 DEGs were detected within the QTL regions previously described in chromosomes 11 and 5, respectively. In addition to these two specific genomic regions, we also observde large transcriptomic changes from genes spread across the genome in the resistant plants in response to the virus infection. This early response against WMV implied genes involved in plant-pathogen interaction, plant hormone signal transduction, the MAPK signaling pathway or ubiquitin mediated proteolysis, in detriment to the photosynthetic and basal metabolites pathways. Moreover, the gene MELO3C021395, which coded a mediator of RNA polymerase II transcription subunit 33A (MED33A), has been proposed as the candidate gene located on chromosome 11 conferring resistance to WMV.ConclusionsThe comparative transcriptomic analysis presented here showed that, even though the resistance to WMV in TGR-1551 has a recessive nature, it triggers an active defense response at a transcriptomic level, which involves broad-spectrum resistance mechanisms. Thus, this study represents a step forward on our understanding of the mechanisms underlaying WMV resistance in melon. In addition, it sheds light into a broader topic on the mechanisms of recessive resistances.

Inheritance of Fusarium wilt resistance in flax

Fusarium wilt (FW) caused by the facultative fungal saprophyte Fusarium oxysporum f. sp. lini (Fol) is a major disease affecting flax production. Cultivation of FW-resistant cultivars mitigates effects of the disease and is a requirement for variety registration. Understanding the genetic inheritance of FW resistance will facilitate the development of resistant varieties. In this study, the inheritance of wilt resistance against western Canadian isolates was studied in F8 recombinant inbred lines (RILs) of a cross between cultivars Aurore (moderately resistant) and Oliver (susceptible). Seventeen Fol isolates were collected from across western Canada and tested for differences in the severity of FW on both parental lines and two check lines under controlled environment conditions. Three of the 17 isolates that caused moderate disease severity were selected to screen the RIL population. The segregation of the RILs for FW resistance was determined under controlled conditions and in field wilt nurseries. A 3:1 ratio of susceptible to resistant RILs was observed with two F. oxysporum f. sp. lini isolates, indicating FW resistance was conditioned by two independent recessive genes. Cluster analysis of disease severity data identified two major clusters of phenotypes in the RIL population, in accordance with the 3S:1 R ratio. High heritability of FW resistance was observed at the late flowering stage, indicating a high level of genotypic variation contributing to phenotypic variation.

Screening of recombinant inbred lines for resistance to bacterial leaf blight pathotypes in rice (Oryza sativa L.)

In the present investigation 16 recombinant inbred lines (RIL’s) developed from the intra-specific cross between YH3 and AKDRMS 21-54 through Marker Assisted Pedigree Breeding Method were screened along with their parents and the checks, namely, BPT 5204, TN1 and Improved Samba Mahsuri (ISM) against IxoPt-20 pathotype at the ICAR-Indian Institute of Rice Research, Hyderabad during Rabi 2021-22 and a new pathotype of Xanthomonas oryzae pv. oryzae causing Bacterial Leaf Blight disease in rice at Regional Agricultural Research Station, Maruteru during Kharif 2022 to identify pathotype specific resistant sources. Morpho-Molecular screening was adopted to evaluate the recombinant inbred lines over two locations in the consecutive seasons of Rabi 2021-22 and Kharif 2022. Based on per cent diseased leaf area, the genotypes were scored and categorised as per the Standard Evaluation System (SES) scale provided by International Rice Research Institute (IRRI). The results revealed all 16 RIL’s to be either resistant (10) or moderately resistant (6) to IxoPt-20 pathotype. However, only five RIL’s were found to be resistant, while four RIL’s were moderately resistant for the new virulent pathotype. Seven RIL’s with resistant to moderately resistant reaction for IxoPt-20 pathotype, showed moderately susceptible reaction for the new virulent pathotype. Among the resistant RIL’s identified for each pathotype, BPT-1901-72-10-6, BPT-1901-108-4-1 and BPT-1901-111-3-2 were found to be uniformly resistant, while, BPT-1901-45-8-6 and BPT-1901-163-1-18 were uniformly moderately resistant to both IXoPt-20 and the new virulent pathotype at Hyderabad and Maruteru, respectively, indicating their potential as genetic stocks for development of new cultivars resistant to bacterial leaf blight disease.

Identification of qBK2.1, a novel QTL controlling rice resistance against Fusarium fujikuroi

BackgroundBakanae disease caused by Fusarium fujikuroi is an increasing threat to rice production. The infected plants show symptoms such as elongation, slenderness, chlorosis, a large leaf angle, and even death. Bakanae disease is traditionally managed by seed treatment. However, fungicide-resistant F. fujikuroi isolates have emerged in several Asian areas, including Taiwan. This study aimed to identify new bakanae resistance quantitative trait loci (QTLs) and provide molecular markers to assist future breeding.ResultsA population of F2:9 recombinant inbred lines (RILs) was derived from the cross between an elite japonica Taiwanese cultivar ‘Taikeng 16 (TK16)’ and an indica variety ‘Budda’. ‘Budda’ was found highly resistant to all 24 representative isolates of the F. fujikuroi population in Taiwan. For the RIL population, 6,492 polymorphic single nucleotide polymorphisms (SNPs) spanning the rice genome were obtained by genotyping-by-sequencing (GBS) technique, and the disease severity index (DSI) was evaluated by inoculation with a highly virulent F. fujikuroi isolate Ff266. Trait-marker association analysis of 166 RILs identified two QTLs in ‘Budda’. qBK2.1 (21.97–30.15 Mb) is a novel and first bakanae resistance QTL identified on chromosome 2. qBK1.8 (5.24–8.66 Mb) partially overlaps with the previously reported qBK1.3 (4.65–8.41 Mb) on chromosome 1. The log of odds (LOD) scores of qBK1.8 and qBK2.1 were 4.75 and 6.13, accounting for 4.9% and 8.1% of the total phenotypic variation, respectively. 64 RILs carrying both qBK1.8 and qBK2.1 showed lower DSI (7%) than the lines carrying only qBK1.8 (15%), only qBK2.1 (13%), or none of the two QTLs (21%). For the future application of identified QTLs, 11 KBioscience competitive allele-specific PCR (KASP) markers and 3 insertion-deletion (InDel) markers were developed.ConclusionsCompared to other important rice diseases, knowledge of bakanae resistance has been insufficient, which limited the development and deployment of resistant cultivars. The discovery of qBK2.1 has provided a new source of bakanae resistance. The resistant RILs inheriting good plant type, good taste, and high yield characteristics from ‘TK16’ can be used as good resistance donors. Our newly developed markers targeting qBK2.1 and qBK1.8 can also serve as an important basis for future fine-mapping and resistance breeding.

Identification of candidate genes associated with resistance against race 0 of Colletotrichum lentis in Lens ervoides

Resistance to anthracnose caused by the fungal pathogen Colletotrichum lentis was explored through transcriptome sequencing over a period of 24 to 96 h post-inoculation (hpi) of the partially resistant recombinant inbred lines (RIL) LR-66-528 and susceptible LR-66-524 of the crop wild relative Lens ervoides population LR-66. The development of infection vesicles and primary hyphae by C. lentis were significantly higher on susceptible RIL LR-66-524 compared to partially resistant LR-66-528 at 24 and 48 hpi, but exponential trends in fungal growth were observed between 24 to 96 hpi in both RILs. Comparison of inoculated with mock-inoculated samples revealed 3091 disease responsive genes, among which 477 were differentially expressed between the two RILs. These were clustered into six expression clusters with genes that had either high or low expression in one of the RILs. Differentially expressed genes (DEGs) were functionally annotated and included genes coding LRR and NB-ARC domain disease resistance proteins, protein detoxification, LRR receptor-like kinase family proteins, and wall-associated Ser/Thr Kinases. DEGs were compared to genes in previously published anthracnose resistance QTLs mapped in LR-66 and revealed 22 DEGs located in 3 QTLs. Expression of 21 DEGs was validated using RT-qPCR confirming expression trends in RNA-seq.

Quantitative Trait Locus Mapping for Fusarium Wilt Race 4 Resistance in a Recombinant Inbred Line Population of Pima Cotton (Gossypium Barbadense).

Fusarium oxysporum f. sp. vasinfectum (FOV) race 4 (FOV4) causes seedling death immediately after emergence, in addition to leaf chlorosis and necrosis, vascular discoloration, plant wilting, defoliation, and plant death at late stages. Breeding for FOV4 resistance is the most cost effective management method. In this study, 163 recombinant inbred lines (RILs) of FOV4-resistant Pima S-6 × susceptible 89590, together with the two parents (Gossypium barbadense), were artificially inoculated with FOV4 and assayed for resistance based on foliar disease severity ratings (DSR) at 30 days post inoculation (dpi) in two replicated tests in the greenhouse or controlled conditions. Significant genotypic variations were detected for FOV4 resistance in a combined analysis of variance. Although a significant genotype × test interaction was detected for DSR, the 10 most resistant RILs had significantly and consistently lower DSR than the susceptible parent in both tests. The heritability estimate for DSR was 0.65, indicating that two-thirds of the phenotypic variation for FOV4 resistance in this Pima RIL population was due to genetic factors. Based on 404 polymorphic SSR markers, five and four quantitative trait loci (QTL) on six chromosomes (c14, c17, c19, c21, c24, and c25) were detected in Tests 1 and 2, respectively, and each explained 15 to 29% of the phenotypic variation. Three QTL on c17, c24, and c25 were in common between the two tests, accounting for 60% and 75% of the QTL detected in Tests 1 and 2, respectively. The three QTL were also reported in previous studies and will be useful for marker-assisted selection for FOV4 resistance in Pima cotton.

Genome-wide characterization of the sorghum JAZ gene family and their responses to phytohormone treatments and aphid infestation

Jasmonate ZIM-domain (JAZ) proteins are the key repressors of the jasmonic acid (JA) signal transduction pathway and play a crucial role in stress-related defense, phytohormone crosstalk and modulation of the growth-defense tradeoff. In this study, the sorghum genome was analyzed through genome-wide comparison and domain scan analysis, which led to the identification of 18 sorghum JAZ (SbJAZ) genes. All SbJAZ proteins possess the conserved TIFY and Jas domains and they formed a phylogenetic tree with five clusters related to the orthologs of other plant species. Similarly, evolutionary analysis indicated the duplication events as a major force of expansion of the SbJAZ genes and there was strong neutral and purifying selection going on. In silico analysis of the promoter region of the SbJAZ genes indicates that SbJAZ5, SbJAZ6, SbJAZ13, SbJAZ16 and SbJAZ17 are rich in stress-related cis-elements. In addition, expression profiling of the SbJAZ genes in response to phytohormones treatment (JA, ET, ABA, GA) and sugarcane aphid (SCA) was performed in two recombinant inbred lines (RILs) of sorghum, resistant (RIL 521) and susceptible (RIL 609) to SCA. Taken together, data generated from phytohormone expression and in silico analysis suggests the putative role of SbJAZ9 in JA-ABA crosstalk and SbJAZ16 in JA-ABA and JA-GA crosstalk to regulate certain physiological processes. Notably, upregulation of SbJAZ1, SbJAZ5, SbJAZ13 and SbJAZ16 in resistant RIL during JA treatment and SCA infestation suggests putative functions in stress-related defense and to balance the plant defense to promote growth. Overall, this report provides valuable insight into the organization and functional characterization of the sorghum JAZ gene family.

Cross-Kingdom Gene Coexpression Analysis UsingaStemphylium botryosum-Lens ervoides System Revealed Plasticity of Intercommunication Between thePathogen Secretome and the Host Immune Systems.

Necrotrophic pathogens are responsible for significant declines in crop yield and quality worldwide. During the infection process, a pathogen releases a series of secretory proteins to counteract the plant immune system, and this interaction of pathogen and host molecules determines whether the pathogen will successfully invade the host plant tissues. In this study, we adopted co-transcriptomic approaches to analyze the Lens ervoides-Stemphylium botryosum system, with a focus on 1,216 fungal genes coding for secretory proteins and 8,810 disease-responsive genes of the host 48, 96, and 144 h postinoculation, captured in two F9 recombinant inbred lines (RILs) displaying contrasting disease responses. By constructing in planta gene coexpression networks (GCNs) for S. botryosum, we found that the pathogen tended to co-upregulate genes regulating cell wall degradation enzymes, effectors, oxidoreductases, and peptidases to a much higher degree in the susceptible host LR-66-577 than in the resistant RIL LR-66-637, indicating that the promotion of these digestive enzymes and toxins increased S. botryosum virulence. Construction of cross-kingdom GCNs between pathogen and plant for the two RILs revealed that the co-upregulation of these fungal digestive enzymes and toxins simultaneously promoted a series of defense responses such as redox change, expression of membrane-related genes and serine/threonine kinase, and stress and disease responses in the susceptible RIL which was not observed in the resistant RIL, indicating that these activities exacerbated susceptibility to S. botryosum.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Evaluation of Resistance to Gummy Stem Blight in a Population of Recombinant Inbred Lines of Watermelon × Citron

Gummy stem blight (GSB), a major disease caused by Stagonosporopsis cucurbitacearum (syn. Didymella bryoniae), has caused significant losses of watermelon in the United States. The lack of progress in the development of resistant cultivars is the result of complex inheritance of resistance and breeding strategies that rely on single-plant selection. Because the sources of resistance are wild watermelon relatives, good fruit quality has been difficult to maintain during the selection process. Three hundred recombinant inbred line (RILs) in a population that carries resistance genes to GSB as well as good fruit quality were produced. This was accomplished by crossing and intercrossing resistant plant introductions, crossing the resulting progenies with elite cultivars, intercrossing those progenies, and, finally, self-pollinating to the S3 generation. The 300 RILs were evaluated for disease severity and fruit morphological and quality traits under greenhouse and field conditions in a randomized complete block design with 10 replications and 3 years. The means and correlations for disease severity ratings and fruit quality traits were estimated. Approximately 186 RILs had disease severity ratings below the mean value of the disease assessment scale (4.5), indicating that they possibly carry one or more genes for resistance to GSB. All disease severity ratings were correlated to each other (r = 0.67–0.98; P < 0.001), but they were not correlated with fruit quality traits. Most importantly, several resistant RILs showed good to excellent fruit quality. Our results provide evidence of improved germplasm with high resistance and good fruit quality.

Introgression of Resistance to Leafminer (Liriomyza cicerina Rondani) from Cicer reticulatum Ladiz. to C. arietinum L. and Relationships between Potential Biochemical Selection Criteria

The chickpea leafminer, Liriomyza cicerina (Rondani), is one of the most destructive insect pests of cultivated chickpea (Cicer arietinum L.) in the Mediterranean region under field conditions. For sustainable and environmentally friendly chickpea production, efforts have been devoted to managing the leafminer via decreasing the use of insecticides. Breeding of new resistant varieties is not only an efficient and practical approach, but also cost-effective and environmentally sensitive. To improve resistant varieties, breeders need reliable biochemical selection criteria that can be used in breeding programs. The first objective was to investigate the possible introgression of resistance to the leafminer from C. reticulatum Ladiz. (resistant) to C. arietinum (susceptible), then, to estimate inheritance of resistance to the leafminer for efficient breeding strategies, and finally, to study organic acid contents as selection criteria. Recombinant inbred lines (RILs) and their parents were evaluated using a visual scale of 1–9 (1 = free from leafminer damage and 9 = mines in more than 91% of the leaflets and defoliation greater than 31%) in the field under natural infestation conditions after the susceptible parent and check had scores of >7 on the visual scale. Superior RILs were found for resistance to the leafminer, and agro-morphological traits indicating that introgression of resistance to leaf miner from C. reticulatum to C. arietinum could be possible using interspecific crosses. The inheritance pattern of resistance to the leafminer in RILs was shown to be quantitative. Organic acids, including oxalic, malic, quinic, tartaric, citric and succinic acids in RILs grown in the field under insect epidemic conditions and in the greenhouse under non-infested conditions were detected by using high performance liquid chromatography (HPLC). In general, organic acids were found to be higher in resistant RILs than susceptible RILs. Path and correlation coefficients showed that succinic acid exhibited the highest direct effects on resistance to the leafminer. Multivariate analyses, including path, correlation and factor analyses suggested that a high level of succinic acid could be used as a potential biochemical selection criterion for resistance to leafminer in chickpea. Resistant RILs with a high seed yield resembling kabuli chickpea can be grown directly in the target environments under leaf miner infestation conditions.

Elucidation of resistance signaling and identification of powdery mildew resistant mapping loci (ClaPMR2) during watermelon-Podosphaera xanthii interaction using RNA-Seq and whole-genome resequencing approach

Watermelon is an important vegetable crop and is widely cultivated in USA with an approximate global production of > 100 million tons. Powdery mildew (PM) caused by Podosphaera xanthii is a major production-limiting factor on watermelon and other cucurbits. Numerous PM and multiple disease resistant (MDR) watermelon germplasm lines have been developed by the USDA in Charleston, SC. To gain a better understanding of the innate and activated molecular defense mechanisms involved during compatible and incompatible PM-watermelon interactions, we inoculated PM susceptible (USVL677-PMS) and resistant (USVL531-MDR) watermelon plants with 105 conidia ml−1 of P. xanthii. RNA-seq profiling was done on leaf samples collected at 0, 1, 3, and 8 days post inoculation (DPI). A total of 2,566 unique differentially expressed genes (DEGs) were identified between compatible and incompatible interactions with P. xanthii. The compatible interactions resulted in distinct plant gene activation (> twofold unique transcripts, 335:191:1762 :: 1:3:8 DPI) as compared to incompatible interaction (> twofold unique transcripts, 314:681:487 :: 1:3:8 DPI). Further, comparative whole-genome resequencing analysis of USVL531-PMR, USVL677-PMS and four introgressed PM resistant recombinant inbred lines (RIL, USVL531-PMR × USVL677-PMS) were performed to identify the region of PM resistance introgressed break points along with other traits inherent by USVL531-PMR by comparing the SNPs and InDels. Based on SNPs identification and CAPS markers, the resistance gene was identified as ClaPMR2, Citrullus lanatus PM Resistance gene 2 {Chr2 : 26750001 .. 26753327 (−)}, a NBS-LRR resistance protein (R) with homology to the Arabidopsis thaliana PM resistance protein, RPW8. The transcriptome data also revealed a complex regulatory network associated with the introgressed junctions mediated by PM resistance R proteins (R genes) that may involve multiple signal regulators and transducers, carbohydrate metabolism, cell wall modifications and the hormone-signaling pathway.

Melon Genome Regions Associated with TGR-1551-Derived Resistance to Cucurbit yellow stunting disorder virus.

Cucurbit yellow stunting disorder virus (CYSDV) is one of the main limiting factors of melon cultivation worldwide. To date, no commercial melon cultivars resistant to CYSDV are available. The African accession TGR-1551 is resistant to CYSDV. Two major quantitative trait loci (QTLs) have been previously reported, both located near each other in chromosome 5. With the objective of further mapping the gene or genes responsible of the resistance, a recombinant inbred line (RIL) population derived from the cross between TGR-1551 and the susceptible cultivar ‘Bola de Oro’ was evaluated for resistance to CYSDV in five different assays and genotyped in a genotyping by sequencing (GBS) analysis. The major effect of one of the two QTLs located on chromosome 5 was confirmed in the multienvironment RIL assay and additionally verified through the analysis of three segregating BC1S1 populations derived from three resistant RILs. Furthermore, progeny test using the offspring of selected BC3 plants allowed the narrowing of the candidate interval to a 700 kb region. The SNP markers identified in this work will be useful in marker-assisted selection in the context of introgression of CYSDV resistance in elite cultivars.

Interactive Gene Expression Patterns of Susceptible and Resistant Lens ervoides Recombinant Inbred Lines and the Necrotroph Ascochyta lentis.

Ascochyta lentis is a foliar pathogen of Lens species and is of worldwide importance in cultivated lentil production. High levels of resistance were identified in the wild species Lens ervoides. This resistance was explored through histopathology, qPCR estimation of fungal biomass and transcriptome sequencing in a susceptible and a resistant recombinant inbred line (RIL) of L. ervoides infected with an aggressive isolate of A. lentis. Necrotrophic growth was delayed in the resistant RIL compared to accelerated necrotrophy of A. lentis in the susceptible RIL. Analysis of the fungal secretome indicated that the early activation of cell wall-degrading enzymes contributed to increased virulence of A. lentis. On the host side, gene co-expression analysis revealed that the invasion by A. lentis caused mRNA, DNA and protein decay in infected plants regardless of the level of resistance in the host. The resistant RIL exhibited a stronger gene co-expression in lipid localization and sulfur processes, and cellular responses to nutrients and stimuli than the susceptible RIL. In addition, differential gene analysis revealed that the repression of both, gibberellin signaling and cell death associated with the hypersensitive response (HR), were associated with enhanced A. lentis resistance.

A Post-Haustorial Defense Mechanism is Mediated by the Powdery Mildew Resistance Gene, PmG3M, Derived from Wild Emmer Wheat.

The destructive wheat powdery mildew disease is caused by the fungal pathogen Blumeria graminis f. sp. tritici (Bgt). PmG3M, derived from wild emmer wheat Triticum dicoccoides accession G305-3M, is a major gene providing a wide-spectrum resistance against Bgt. PmG3M was previously mapped to wheat chromosome 6B using an F6 recombinant inbred line (RIL) mapping population generated by crossing G305-3M with the susceptible T. durum wheat cultivar Langdon (LDN). In the current study, we aimed to explore the defense mechanisms conferred by PmG3M against Bgt. Histopathology of fungal development was characterized in artificially inoculated leaves of G305-3M, LDN, and homozygous RILs using fluorescence and light microscopy. G305-3M exhibited H2O2 accumulation typical of a hypersensitive response, which resulted in programmed cell death (PCD) in Bgt-penetrated epidermal cells, while LDN showed well-developed colonies without PCD. In addition, we observed a post-haustorial resistance mechanism that arrested the development of fungal feeding structures and pathogen growth in both G305-3M and resistant RIL, while LDN and a susceptible RIL displayed fully developed digitated haustoria and massive accumulation of fungal biomass. In contrast, both G305-3M and LDN exhibited callose deposition in attempt to prevent fungal invasion, supporting this as a mechanism of a basal defense response not associated with PmG3M resistance mechanism per se. The presented results shed light on the resistance mechanisms conferred by PmG3M against wheat powdery mildew.

Genetic mapping of angular leaf spot resistance to Pseudomonas syringae pv. lachrymans in a Cucumis hystrix introgression line of cucumber

Angular leaf spot (ALS), caused by Pseudomonas syringae pv. lachrymans, is a common and devastating bacterial disease of cucumber worldwide. However, to the best of our knowledge, the report on genetic mapping of ALS resistance in cucumber is relatively few. In this study, IL52, a Cucumis hystrix introgression line of cucumber, was identified to be resistant to ALS. To investigate the inheritance and genomic location of the ALS resistance gene, IL52 was crossed with a susceptible parent changchunmici (CCMC) to develop F1 plant and 155 F6 recombinant inbred lines (RILs). The F1 plant was susceptible to ALS. The segregation ratio of ALS disease resistance in 155 RILs was 80:75 resistance/susceptibility, fitting to 1:1. The results indicated that a single recessive locus psl-1 was responsible for conferring resistance to ALS in IL52. Further, psl-1 was mapped into the 3,118,855–3,951,558 bp interval of chromosome 1 by combining molecular mapping with bulked segregant analysis and next-generation sequencing (BSA-seq). The marker ALS-InDel was found to be closely linked with psl-1 through the genetic linkage analysis in the 18 resistant RILs, 18 susceptible RILs and 19 susceptible cucumber cultivars. This study on genetic mapping of ALS resistance gene in a Cucumis hystrix introgression line of cucumber will be helpful for fine mapping and marker-assisted selection of ALS resistance gene.

Comparison of the penetration, development and reproduction of Meloidogyne graminicola, and analysis of lignin and total phenolic content in partially resistant and resistant recombinant inbred lines of Oryza sativa

The mechanism of resistance of selected recombinant Oryza sativa inbred lines (RILs) resistant and partially resistant to Meloidogyne graminicola, and their resistant (IR78877–208-B-1-2) and susceptible (IR64, Dinorado) parents was examined by comparing the penetration and development of the second-stage juveniles (J2) and females with the nematode infection process in the resistant Oryza glaberrima genotype TOG5674. Lignification and total phenolic content of infected and uninfected plants were also studied. Less J2 were able to penetrate the roots of the partially resistant and resistant O. sativa genotypes included in the study. The development of J2 that had penetrated in the roots of the RILs was delayed resulting in turn in a delay in the development of egg-laying females and the second generation. Lignification was more intense in infected plants compared with uninfected plants and in the resistant genotypes (especially TOG5674) compared with the susceptible genotypes. This observation suggests that lignification, at least in some rice genotypes, is constitutively expressed and that this expression may differ in time. The accumulation of lignin in the epidermis of some RILs and the endodermis of TOG5674 at 1 day after inoculation suggests that lignification may act as a constitutive post-penetration mechanism of resistance to M. graminicola infection at least in some rice genotypes. On the other hand, higher intensity of lignification in resistant vs susceptible genotypes and the accumulation of lignin in the endodermis at the site of giant cell formation may suggest that lignification also acts as an inducible post-penetration mechanism of resistance to M. graminicola infection. A higher total phenolic content was observed in infected plants of some RILs compared with uninfected plants during the first weeks of observation.

Antioxidants elevates the resistance to Cercospora canescens in interspecific cross of Vigna radiata (Kopergaon) × Vigna mungo (Pant Urd 31)

Cercospora leaf spot (CLS) caused by Cercospora canescens is one of the most important foliar diseases responsible for 30%–75% yield losses in mungbean. CLS is induced by the photo-activated toxin cercosporin. The most operative measure of managing the disease is the use of resistant varieties. The objective of present study was to determine the changes in the antioxidants status of mung-bean plant to differential responses of Cercospora leaf spot under field conditions. The present study attempted to quantify the antioxidant activities against C. canescens infection in resistant RILs of inter specific cross of Vigna radiata (Kopergaon) × Vigna mungo (PU31). It was found that antioxidants such as superoxide dismutase (SOD) and catalase (CAT) enzymes were actively involved in scavenging ROS in both resistant and susceptible recombinant inbred lines (RILs) after CLS infection. Throughout the infection, the activity of both the enzymes were higher in resistant than in susceptible RILs. The malondialdehyde (MDA) accumulated at a significantly low concentration in resistant RILs and higher in susceptible ones. Statistically, area under disease progress curve (AUDPC) and MDA showed a significant positive correlation whereas, there was a negative correlation between AUDPC and SOD and CAT activity. The disease components such as lesion number and total lesion size were variable in RILs with different disease reactions. The study identified few RILs resistant to Cercospora leaf spot infection in response to utilization of the antioxidant system to confer Cercospora leaf spot resistance in mungbean by detoxifying cercosporin induced reactive oxygen species.

English

Parasitic weed Striga gesnerioides (Willd.) is one of the major constraints of cowpea production. Host-plant resistance seems to be efficient and economical in controlling the pest. The objectives of this study were to evaluate recombinant inbred lines developed between IT97K 499-35 (Striga resistant parent,) and Sanzi (susceptible parent), by Single Seed Descent (SSD), for Striga resistance in Northern Ghana. The study also evaluated the promising Striga gesnerioides resistant lines and susceptible checks for yield loss due to Striga infestation. The studies involved a field and pot screening under artificial inoculation. Twenty-seven (27) recombinant inbred lines (RILs) out of the 251 RILs screened were resistant to Striga gesnerioides. The percentage reduction in the grain yield and dry biomass were lower in the resistant RILs (0.55 to 3.08% and 1.11 to 7.7%, respectively) than the susceptible ones (28.45 to 58.88% and 47.29 to 61.71%, respectively). The negative effect of Striga infestation on cowpea grain yield and dry biomass can then be reduced when resistant genotypes are used. Key words: Cowpea, Striga gesnerioides, recombinant inbred lines, yield loss.