Susceptible Commercial Cultivar Research Articles

Evaluation of Endophytic Microorganisms (Trichoderma hazianum and Bacillus subtilus) as Biofungicides against Bayoud Disease (Fusarium oxysporum f. sp. albedinis) on Date Palm (Phoenix dactylifera L.)

Backgraound: Date palm disease named Bayoud, is the major constraint caused by the soil fungus Fusarium oxysporum f. sp. albedinis (Foa) in Algeria. This work has the objective to in vivo biocontrol on bayoud disease incidence on three date palm varieties (Deglet-Nour, Feggous and H-mira), using two antagonistic microorganisms, one fungus (Trichoderma harzianum) and one bacterium (Bacillus subtilus). Methods: Foa strains obtained from isolation of the spines carrying the typical symptoms of Bayoud disease. Two strains used T. harzianum and B. subtilus, are isolated from rhizosphere soils of the date palm. Using the greenhouse screening test, the in vivo biocontrol test is carried out on 3-4 month-old date palm seedlings at the rate of 5 ml of inoculum suspension. Result: A significant (P less than 0.01), difference of date palm seedlings reactions against Foa isolate and antagonistic microorganisms was observed. All three date palm varieties present a high susceptibility against Foa until the total mortality of date palm seedlings. The biocontrol test showed that two antagonists showed different reaction, with the complete resistance of three varieties treated by T. harzianum, while, the antagonist B. subtilus protect just two varieties (Deglet-Nour and H-mira) and can’t protect Feggous variety when showed the susceptibility reaction by presentation of typical symptoms of bayoud disease. We can apply of these antagonistic microorganisms to protect our groves contaminated by Bayoud disease and contain just susceptible commercial cultivars of date palm.

Survival of Intervarietal Grafts of Sweet and Serrano Peppers (Capsicum annuum L.) on Pasilla 18M as a New Rootstock Against Phytophthora capsici Leo

The genus Capsicum has been distinguished by its lack of compatible rootstocks with commercial cultivars to successfully protect against Phytophthora capsici. Criollo de Morelos 334 (CM334) has been used worldwide in crosses and as a rootstock to protect against P. capsici. However, novel sources of resistance to this pathogen, such as ‘Pasilla 18M’ have not yet been explored as rootstocks. A good rootstock should be highly compatible with the scion and also maintain the quality and/or provide a benefit to the grafted cultivar. Our objectives were 1) to evaluate grafting survival using ‘Pasilla 18M’ and CM334 as rootstocks of two susceptible commercial cultivars: Sweet Pepper California Wonder (CW) and Serrano Coloso; and 2) to evaluate the efficiency of ‘Pasilla 18M’ as rootstock against P. capsici using CM334 as a resistant control. Grafting survival was analyzed over 58 days after grafting in sets of 60 plants per varietal combination. Disease severity and incidence were recorded during 24 days after inoculation with P. capsici (DAI). Incidence was also evaluated at 54 and 84 DAI. A severity scale from 0 (healthy plant) to 4 (dead plant) was applied to evaluate root rot per plant. Incidence was recorded as the percent of diseased plants (severity >0). Grafting survival of intervarietal grafts was 87% to 94%, similar to ungrafted cultivars, and exceeding autograft survival. Ungrafted and autografted Sweet Pepper and Serrano showed root rot severities 2.3 to 3.3, with 89% to 100% incidence. In contrast, intervarietal grafts remained almost free of infection (severity 0.14; incidence 0% to 4%). CM334 and ‘Pasilla 18M’ rootstocks are highly compatible with ‘Serrano Coloso’ and ‘Sweet Pepper CW’. ‘Pasilla 18M’ confers the same level of protection against P. capsici as CM334.

Evaluation of pea genotype PI180693 partial resistance towards aphanomyces root rot in commercial pea breeding.

The cultivation of vining pea (Pisum sativum) faces a major constraint with root rot diseases, caused by a complex of soil-borne pathogens including the oomycetes Aphanomyces euteiches and Phytophtora pisi. Disease resistant commercial varieties are lacking but the landrace PI180693 is used as a source of partial resistance in ongoing pea breeding programs. In this study, the level of resistance and their interaction with A. euteiches virulence levels of six new back-crossed pea breeding lines, deriving from the cross between the susceptible commercial cultivar Linnea and PI180693, were evaluated for their resistance towards aphanomyces root rot in growth chamber and green house tests. Resistance towards mixed infections by A. euteiches and P. pisi and commercial production traits were evaluated in field trials. In growth chamber trials, pathogen virulence levels had a significant effect on plant resistance, as resistance was more consistent against A. euteiches strains exhibiting high or intermediate virulence compared with lowly virulent strains. In fact, line Z1701-1 showed to be significantly more resistant than both parents when inoculated with a lowly virulent strain. In two separate field trials in 2020, all six breeding lines performed equally well as the resistant parent PI180693 at sites only containing A. euteiches, as there were no differences in disease index. In mixed infections, PI180693 exhibited significantly lower disease index scores than Linnea. However, breeding lines displayed higher disease index scores compared with PI180693, indicating higher susceptibility towards P. pisi. Data on seedling emergence from the same field trials suggested that PI180693 was particularly sensitive towards seed decay/damping off disease caused by P. pisi. Furthermore, the breeding lines performed equally well as Linnea in traits important for green pea production, again emphasizing the commercial potential. In summary, we show that the resistance from PI180693 interacts with virulence levels of the pathogen A. euteiches and is less effective towards root rot caused by P. pisi. Our results show the potential use of combining PI180693 partial resistance against aphanomyces root rot with commercially favorable breeding traits in commercial breeding programs.

Characterisation of an ethyl methanesulfonate‐derived drought‐tolerant sugarcane mutant line

AbstractSustainable sugarcane production in areas prone to frequent and severe drought can be achieved by creating resilient sugarcane varieties. In this study, a unique mutant line, M9.2, was generated from a drought susceptible commercial sugarcane cultivar, N19, through the exposure of callus cells to the ethyl methanesulfonate mutagen and subsequent in vitro osmotic selection on polyethylene glycol. The study aimed to characterise the M9.2 mutant, in comparison with the parental genotype, in terms of its physiological and biochemical performance and proteome profile when exposed to moderate (14 days without water) and severe (21 days without water) water deficit stress in glasshouse pot trials. In comparison to the parental counterparts, the mutant plants were able to sustain the quantum efficiency of photosystem II (Fv/Fm) throughout the stress. Under mild stress, the mutant plants displayed elevated stomatal conductance, high concentrations of proline, accumulated less H2O2 and phenotypically displayed limited wilting and no visible signs of leaf senescence. Under severe stress, the mutant plants accumulated less malondialdehyde and more antioxidant enzymes (superoxide dismutase and catalase) than the parental line. Differential protein expression was also observed according to two‐dimensional difference gel electrophoresis patterns of proteins expressed in the M9.2 mutant versus the parental plants during moderate stress. Analysis revealed proteins related to photosynthesis (pyruvate orthophosphate dikinase, un‐fragmented ribulose‐1,5‐bisphosphate carboxylase/oxygenase large subunit and chlorophyll a/b‐binding protein 3) and carbohydrate metabolism (sucrose synthase) were up‐regulated in the mutant. Differentially expressed proteins were further linked to energy metabolism, methylation homeostasis and DNA repair. This study characterises the new M9.2 mutant with beneficial drought‐tolerant traits which have the potential to be exploited in future sugarcane breeding programmes.

Gene Expression, Histology and Histochemistry in the Interaction between Musa sp. and Pseudocercospora fijiensis.

Bananas are the main fruits responsible for feeding more than 500 million people in tropical and subtropical countries. Black Sigatoka, caused by the fungus Pseudocercospora fijiensis, is one of the most destructive disease for the crop. This fungus is mainly controlled with the use of fungicides; however, in addition to being harmful to human health, they are associated with a high cost. The development of resistant cultivars through crosses of susceptible commercial cultivars is one of the main focuses of banana breeding programs worldwide. Thus, the objective of the present study was to investigate the interaction between Musa sp. and P. fijiensis through the relative expression of candidate genes involved in the defence response to black Sigatoka in four contrasting genotypes (resistant: Calcutta 4 and Krasan Saichon; susceptible: Grand Naine and Akondro Mainty) using quantitative real-time PCR (RT–qPCR) in addition to histological and histochemical analyses to verify the defence mechanisms activated during the interaction. Differentially expressed genes (DEGs) related to the jasmonic acid and ethylene signalling pathway, GDSL-like lipases and pathogenesis-related proteins (PR-4), were identified. The number and distance between stomata were directly related to the resistance/susceptibility of each genotype. Histochemical tests showed the production of phenolic compounds and callosis as defence mechanisms activated by the resistant genotypes during the interaction process. Scanning electron microscopy (SEM) showed pathogenic structures on the leaf surface in addition to calcium oxalate crystals. The resistant genotype Krasan Saichon stood out in the analyses and has potential for use in breeding programs for resistance to black Sigatoka in banana and plantains.

Potato plants transformed with the Arabidopsis EF-Tu receptor (EFR) show restricted pathogen colonization and enhanced bacterial wilt resistance under conditions resembling natural field infections

Potato is considered a staple food in the world and its production is limited by the presence of bacterial wilt (bw) disease caused by Ralstonia solanacearum. Host resistance is the most sustainable and cost-effective strategy to manage bw, although resistant commercial potato cultivars are not yet available. Our group incorporated the efr receptor of Arabidopsis thaliana (atefr), which recognizes the elongation factor Tu, preserved in bacteria, triggering an immune response. atefr was tested in two genetic backgrounds: a susceptible commercial cultivar (inia Iporá) and a breeding clone with partial resistance introgressed from Solanum commersonii. In this work, the effect of the atefr receptor on bw resistance was evaluated for the first time, using conditions resembling natural field infection. In addition, the colonization patterns of wild-type and transgenic lines were compared by using luminescent and fluorescent R. solanacearum reporter strains. Both approaches showed a delay and a decrease in the severity of wilting symptoms in the atefr-transformed genotypes. Differential colonization patterns were observed, revealing a higher bacterial development in the non-transformed plants. This atefr effect seems more pronounced in the interspecific breeding line, possibly leading to a more effective activation of the plant immune system.

Genetic diversity of Puccinia kuehnii, the causal agent of orange rust of sugarcane, from Brazil

AbstractThe use of resistant genotypes is the preferred method to control orange rust of sugarcane (Saccharum spp) caused by Puccinia kuehnii. This approach has been adopted in Brazil but outbreaks of the disease on previously resistant varieties showed that the efficacy of this method is limited and requires a better understanding of pathogen diversity. Nevertheless, adequate molecular markers for examining pathogen diversity at population level are not available, which limits the success of orange rust control by genetic resistance. Therefore, two independent investigations were conducted to examine genetic diversity of P. kuehnii from São Paulo state, the most important sugarcane growing state of Brazil. First, simple‐sequence repeat (SSR) markers were developed in the present work and genotypic diversity of orange rust isolates from different locations investigated. Second, phenotypic diversity was examined by the single‐pustule inoculation technique on P. kuehnii isolates retrieved from three susceptible commercial sugarcane cultivars. A total of 96 SSR markers were generated and tested for this species. Subsequently, 29 isolates of P. kuehnii were fingerprinted with nine SSR markers to estimate the genotypic diversity by neighbour‐joining and 3D principal coordinates. The 29 isolates of the pathogen clustered into four main groups, which were identified by three SSR markers (NPRL_PK_108a, NPRL_PK_162_spka and NPRL_PK_221_spka). Phenotypic data at 21 days after the single‐pustule inoculation showed that P. kuehnii from highly susceptible commercial cultivars harboured a small proportion of variants capable of causing disease on resistant cultivars. A differential reaction was demonstrated for the most virulent variant in a repeated experiment confirming the existence of races within P. kuehnii in Brazil.

Identification of NBS-LRR Resistance Gene Analogues (RGA) from Rose (IIHRR13-4) Resistant to Powdery Mildew (Podosphaera pannosa (Wallr.:Fr.) de Bary)

Resistance is the best strategy to manage powdery mildew (Podosphaera pannosa (Wallr.:Fr.) de Bary) of rose. Identification of resistant genes (R genes) from plant species will help in breeding programs. Nucleotide Binding Site - Leucine Rich Repeats (NBS- LRR) is a major class of R gene family in plants. This study reports the identification and molecular characterization of resistance gene analogues from roses maintained at ICAR- Indian Institute of Horticultural Research (IIHR). The powdery mildew resistant line IIHRR13-4 was compared with the susceptible commercial cultivar, konfetti. PCR based approaches with degenerative primers based on different conserved motifs of NBS-LRR were employed to isolate resistance gene analogues (RGAs) from rose. Eleven RGAs (IIHRR13-4R1, IIHRR13-4R2, IIHRR13-4R3, IIHRR13-4R4, IIHRR13-4R5, IIHRR13- 4R6, IIHRR13-4R7, IIHRR13-4R8 IIHRR13-4R9 and IIHRR13-4R10) were identified from powdery mildew resistant germplasm line, IIHRR13-4, based on the sequence and similarity to RGAs from rosaceae family and other crops. The major similarity to rose RGAs reported are from Fragaria vesca, Rosa hybrid cultivar, Prunus and Rosa chinensis. RGAs isolated from IIHRR13-4 belonged to Toll Interleukin Receptor (TIR)-NBS-LRR and Non-TIR-NBS-LRR RGAs (Lecine Zipper (LZ) type). Different motifs of RGAs identified were P-loop, RNBS A, kinase 2, kinase 3a, RNBS-D and GLPL of NBS domain. This study reports the existence of resistance at genetic level in powdery mildew resistant genotype IIHRR13-4. These RGAs will be useful for mapping and characterization of R genes in IIHRR13-4 and breeding for improved powdery mildew resistance in roses.

Identification of NBS-LRR Resistance Gene Analogues (RGA) from Rose (IIHRR13-4) Resistant to Powdery Mildew (Podosphaera pannosa (Wallr.:Fr.) de Bary)

Resistance is the best strategy to manage powdery mildew (Podosphaera pannosa (Wallr.:Fr.) de Bary) of rose. Identification of resistant genes (R genes) from plant species will help in breeding programs. Nucleotide Binding Site - Leucine Rich Repeats (NBS- LRR) is a major class of R gene family in plants. This study reports the identification and molecular characterization of resistance gene analogues from roses maintained at ICAR- Indian Institute of Horticultural Research (IIHR). The powdery mildew resistant line IIHRR13-4 was compared with the susceptible commercial cultivar, konfetti. PCR based approaches with degenerative primers based on different conserved motifs of NBS-LRR were employed to isolate resistance gene analogues (RGAs) from rose. Eleven RGAs (IIHRR13-4R1, IIHRR13-4R2, IIHRR13-4R3, IIHRR13-4R4, IIHRR13-4R5, IIHRR13- 4R6, IIHRR13-4R7, IIHRR13-4R8 IIHRR13-4R9 and IIHRR13-4R10) were identified from powdery mildew resistant germplasm line, IIHRR13-4, based on the sequence and similarity to RGAs from rosaceae family and other crops. The major similarity to rose RGAs reported are from Fragaria vesca, Rosa hybrid cultivar, Prunus and Rosa chinensis. RGAs isolated from IIHRR13-4 belonged to Toll Interleukin Receptor (TIR)-NBS-LRR and Non-TIR-NBS-LRR RGAs (Lecine Zipper (LZ) type). Different motifs of RGAs identified were P-loop, RNBS A, kinase 2, kinase 3a, RNBS-D and GLPL of NBS domain. This study reports the existence of resistance at genetic level in powdery mildew resistant genotype IIHRR13-4. These RGAs will be useful for mapping and characterization of R genes in IIHRR13-4 and breeding for improved powdery mildew resistance in roses.

The effects of Meloidogyne incognita and Heterodera glycines on the yield and quality of edamame (Glycine max l.) in Arkansas.

In 2012, the first domestic commercial edamame processing plant was established in Arkansas and edamame production was contracted out to local growers. Although the state is a major soybean producer, studies of nematode effects on edamame are limited. A survey of nematode genera and density in 64 contracted edamame production fields was conducted in 2013 and 2014. In both years, Meloidogyne and Heterodera were present in less than half of the surveyed fields while Pratylenchus was the most prevalent in 2013 and Helicotylenchus in 2014. A microplot study was conducted in 2014 in two locations to evaluate the effects of root-knot nematode (Meloidogyne incognita, race 3) and soybean cyst nematode (Heterodera glycines, HG type 2.5.7) on plant growth, yield and food quality components of edamame. Yield was the most consistent factor influenced by nematode pressure with increasing nematode population densities resulting in suppressed pod and seed weight. Additionally, seed protein content was reduced in the highest tested population density of H. glycines. In greenhouse studies, 22 advanced edamame breeding lines from the University of Arkansas soybean breeding program were compared with two susceptible commercial cultivars for suitability as hosts for both M. incognita and H. glycines independently. Four lines showed consistent reductions in M. incognita reproduction relative to the commercial cultivars and could represent sources of moderate resistance for development of future root-knot nematode resistant edamame cultivars.

Feasibility of omega bench grafting in guava tree (Psidium guajava L.) propagated via herbaceous stem cutting

ABSTRACTRoot-knot nematode Meloidogyne mayaguensis Rammah & Hirschmann is a limiting factor in guava tree (Psidium guajava L.) production around the world. An alternative to address this problem is the use of resistant root-knot nematode rootstocks. However, the guava nursery system which is based on herbaceous stem cutting propagation needs to be modified as a rootstock produced via cutting must be grafted with nematode susceptible commercial cultivars. Thus, the objective of this study was to verify the feasibility of omega bench grafting in guava tree propagated by herbaceous stem cutting. Root-knot nematode tolerant ‘Tailandesa’ herbaceous stem cuttings was used as rootstock and ‘Paluma’ and ‘Pedro Sato’ as commercial cultivar grafts. It was observed 100% rooting, cutting survival, and leaf retention on rootstocks grafted with ‘Paluma’ and ‘Pedro Sato’. However, grafting take was not successful as the tissues presented fast oxidation, dehydration and graft death in both commercial cultivars. However, the immersion of the grafts into a solution containing 2.0% citric acid and 1.0% ascorbic acid controlled the oxidation process and resulted in 28.6% and 37.5% successful grafting in the combinations ‘Paluma’/‘Tailandesa’ and ‘Pedro Sato’/’Tailandesa’, respectively.

A fitness cost resulting from Hamiltonella defensa infection is associated with altered probing and feeding behaviour in Rhopalosiphum padi.

Many herbivorous arthropods, including aphids, frequently associate with facultative endosymbiotic bacteria, which influence arthropod physiology and fitness. In aphids, endosymbionts can increase resistance against natural enemies, enhance aphid virulence and alter aphid fitness. Here, we used the electrical penetration graph technique to uncover physiological processes at the insect-plant interface affected by endosymbiont infection. We monitored the feeding and probing behaviour of four independent clonal lines of the cereal-feeding aphid Rhopalosiphum padi derived from the same multilocus genotype containing differential infection (+/-) with a common facultative endosymbiont, Hamiltonella defensa Aphid feeding was examined on a partially resistant wild relative of barley known to impair aphid fitness and a susceptible commercial barley cultivar. Compared with uninfected aphids, endosymbiont-infected aphids on both plant species exhibited a twofold increase in the number of plant cell punctures, a 50% reduction in the duration of each cellular puncture and a twofold higher probability of achieving sustained phloem ingestion. Feeding behaviour was also altered by host plant identity: endosymbiont-infected aphids spent less time probing plant tissue, required twice as many probes to reach the phloem and showed a 44% reduction in phloem ingestion when feeding on the wild barley relative compared with the susceptible commercial cultivar. Reduced feeding success could explain the 22% reduction in growth of H. defensa-infected aphids measured on the wild barley relative. This study provides the first demonstration of mechanisms at the aphid-plant interface contributing to physiological effects of endosymbiont infection on aphid fitness, through altered feeding processes on different quality host plants.

The Effect of Pathogenesis-Related 10 (PR-10) Gene on the Progression of Fusarium Wilt in Musa acuminata cv. Berangan

PR-10 is a member of pathogenesis-related (PR) genes elicited by the plant’s defense mechanism during pathogen attack. Elevated expression of PR-10 upon different pathogen invasions has been observed in many plant species suggesting its role as an anti-bacterial, anti-viral and anti-fungal gene. However, the effect of PR-10 in mitigating the infection of Fusarium oxysporum f. sp. cubense (Foc), the causal agent of Fusarium wilt in banana has not been reported. In this study, the coding sequences of PR-10 gene isolated from Foc resistant Musa acuminata ssp. malaccensis (MaPR-10) were integrated into a local Foc susceptible commercial banana cultivar, Berangan via co-cultivation of embryogenic cell suspension and Agrobacterium tumefaciens. Out of 17 putative transgenic lines established, 11 of them positively harbored MaPR-10. Among these, Line-19 plantlets showed the most rapid in-vitro propagation and successfully over-expressed the transgene. Following a nursery challenge experiment with a virulent Foc race 4 (CI HIR) isolate, about 30% of Line-19 plants showed a one-week delay in disease progression when compared to the untransformed controls. From the final evaluation performed in the 5th week-post-inoculation, the leaf symptoms index (LSI) and rhizome discoloration index (RDI) of Line-19 was 3.4 and 6.1, respectively, indicating the disease had progressed. The findings of this study enrich the current existing knowledge on the roles of PR-10 in combating fungal disease in plants.

High-resolution genetic mapping of a novel brown planthopper resistance locus, Bph34 in Oryza sativa L. X Oryza nivara (Sharma & Shastry) derived interspecific F2 population.

A BPH-resistant locus designated as Bph34 identified in Oryza nivara acc. IRGC104646 on long arm of chromosome 4 using high-resolution mapping with 50K SNP chip. BPH resistance contributed by locus showed dominant inheritance in F2 and F3. The Bph34 locus is 91kb in size and contains 11 candidate genes. In addition to SNP markers, SSR markers, RM16994 and RM17007 co-segregated with the BPH resistance. These two SSR markers can facilitate marker-assisted transfer of the Bph34 locus into elite rice cultivars in all labs. Brown planthopper (BPH, Nilaparvata lugen Stål) is one of the most destructive insects of rice (Oryza sativa L.) causing significant yield losses annually. Exploiting host plant resistance to BPH and incorporating resistant genes in susceptible commercial cultivars is economical and environmentally friendly approach to manage this pest. Here, we report high-resolution mapping of a novel genetic locus for resistance to BPH, designated as Bph34 on long arm of rice chromosome 4. The locus was mapped using an interspecific F2 population derived from a cross between susceptible indica cultivar PR122 and BPH-resistant wild species, O. nivara acc. IRGC104646. Inheritance studies performed using F2 and F2:3 populations revealed the presence of single dominant gene. Construction of high-density linkage map using 50K SNP chip (OsSNPnks) followed by QTL mapping identified single major locus at 28.8 LOD score between SNP markers, AX-95952039 and AX-95921548. The major locus contributing resistance to BPH designated as Bph34 and explained 68.3% of total phenotypic variance. The Bph34 locus is 91Kb in size on Nipponbare reference genome-IRGSP-1.0 and contains 11 candidate genes. In addition to associated SNP markers, two SSR markers, RM16994 and RM17007, also co-segregated with the Bph34 which can be used efficiently for markers assisted transfer into elite rice cultivars across the labs.

A Rapid Phenotyping Method for Imazamox Resistance in Wheat

ABSTRACT: The availability of imidazolinone (IMI) resistant cultivars has provided an effective option for weed control in wheat production systems. IMI herbicides control several weeds, including Avena fatua and Lolium multiflorum, which are the most frequent grass weeds in wheat crops of the Argentine Pampas. The aim of this study was to develop a soil-less method that allows rapid phenotyping of IMI resistance in wheat. Nine wheat cultivars differing in IMI resistance were evaluated through a between-paper germination method. Herbicide concentrations required to reduce shoot and root length by 50% for resistant cultivars were > 75 fold that of the susceptible cultivars. The response of resistant and susceptible commercial cultivars was assessed in the between-paper and top-paper methods at 100 mM imazamox as discriminating dose. The Z’-factor was calculated for evaluation of the quality of the screening methods. Both germination methods showed Z’-factors > 0 indicating that the assays were appropriate but the between-paper method allowed to save space in the growth chamber. The germination methods were useful for distinguishing between susceptible and resistant plants carrying at least one resistance gene. The rapid, simple and cost-effective method described in the present study could be a potential tool when selecting for IMI resistance in wheat in breeding programs.

Characterization of host plant resistance to zebra chip disease from species-derived potato genotypes and the identification of new sources of zebra chip resistance.

‘Candidatus Liberibacter solanacearum’ (Lso), an uncultivable phloem-limited phytopathogenic bacteria, is known to be associated with Zebra Chip disease (ZC), which represents a major threat to potato production in the US and elsewhere. This pathogen is transmitted by the phloem-feeding potato psyllid, Bactericera cockerelli Sulc (Hem. Triozidae). Currently, there are no reports of resistance to ZC in cultivated potatoes. This greenhouse study was conducted to evaluate Lso transmission success and the susceptibility of 11 potato breeding clones, representing diverge genetic background, to ZC, in relation to a susceptible commercial cultivar, Russet Burbank. Individual plants were exposed to two Lso-positive potato psyllids for 48 hours. The percentage of successful Lso transmission varied across the evaluated genotypes ranging between 7 and 57%. Freshly-cut and fried tubers showed significant variation in ZC symptom severity among the breeding clones, with several genotypes expressing relative tolerance when compared to Russet Burbank. None of the evaluated clones showed statistically lower Lso titers than Russet Burbank with the exception of one genotype in the second year of the study. However, the presence of a non-significant relationship between average symptom severity and Lso titer indicated variations in phenotypic responses (i.e. tolerance) to Lso existed among evaluated breeding lines. Breeding clones A07781-3LB, A07781-4LB and A07781-10LB had relatively lower Lso titer (low susceptibility) and tuber symptom expression (high tolerance) among the tested genotypes. As these three clones represent full siblings, the observed effects could be indicative of the presence of a genetic basis for resistance/tolerance to ZC. Findings provide a better understanding of resistance/tolerance to ZC, and contribute to continued efforts in breeding for resistance to this disease.

Marker-Assisted Development and Evaluation of Near-Isogenic Lines for Broad-Spectrum Powdery Mildew Resistance Gene Pm2b Introgressed into Different Genetic Backgrounds of Wheat.

At present, most of released wheat cultivars or breeding lines in China are susceptible to powdery mildew (Pm) (caused by Blumeria graminis f. sp. tritici, Bgt), so there is an urgent need to rapidly transfer effective and broad-spectrum Pm resistance genes into elite cultivars/lines. Near-isogenic lines (NILs) with short target gene region are very important in molecular breeding and map-based cloning and can be developed by combining marker-assisted selection and conventional phenotypic identification. However, no Pm gene NILs were reported by using this method in the previous studies. A new broad-spectrum dominant resistance gene Pm2b, derived from the Chinese wheat breeding line KM2939, conferred high resistance to Pm at both the seedling and adult stages. In this study, with the aid of forward and background selection (FS and BS) using molecular markers, the Pm2b gene was introgressed into three elite susceptible commercial cultivars Shimai 15, Shixin 828, and Kenong 199 through the back-crossing procedure. With the appropriate backcrossing generations, selected population sizes and marker number for BS, the homozygous resistant BC3F2:3 NILs of Pm2b gene in the three genetic backgrounds with the highest recipient genome composition of about 99%, confirmed by simple sequence repeat markers and 660K single nucleotide polymorphic array, were developed and evaluated for the powdery mildew resistance and agronomic traits. The different resistance and similar or improved agronomic performance between Pm2b NILs and their corresponding recurrent parents indicated their potential value in the marker-assisted breeding of the Pm2b gene. Moreover, the development of four flanked diagnostic markers (CFD81, BWM25, BWM20, and BWM21) of the Pm2 gene can effectively assist the forward selection and accelerate the transfer and use of this resistance gene.

GENETIC ANALYSIS OF RESISTANCE TO OROBANCHE CRENATA (FORSK.) IN A PEA (PISUM SATIVUM L.) LOW-STRIGOLACTONE LINE

Crenate broomrape ( Orobanche crenata Forsk.) is a root parasitic weed severely threatening the cultivation of pea ( Pisum sativum L.) in Mediterranean agricultural settings. We recently characterized a pea line highly resistant to O . crenata , named ROR12. Analysis of root exudates indi- cated that ROR12 resistance is possibly due to lower exu- dation of strigolactones, carotenoid-derived compounds acting in the rhyzosphere as broomrape germination stim- ulants. Here, we provide a first genetic characterization of ROR12 resistance. Two segregating F 2 populations origi- nating from the cross between ROR12 and two susceptible commercial pea cultivars were assayed for response to O . crenata , together with the respective parental lines and F 1 generations. Susceptibility of F 1 individuals indicated that resistance is recessively inherited. In F 2, significant devia- tion from Gaussian distribution suggested that broomrape resistance is not a quantitative trait, as it was previously hypothesized, but is rather controlled by one or a few ma- jor loci. Molecular analysis with RAPD and SSR markers identified a genomic region on pea linkage group 3 associ- ated with the resistant phenotype. Finally, transcriptional evidence was provided, supporting the hypothesis that ROR12 resistance is due to reduced strigolactone biosyn- thesis. Overall, this study provides insights on the control of broomrape resistance and lays the foundation for future work addressing the fine-mapping and cloning of genes controlling strigolactone biosynthesis and/or broomrape resistance.

Metabolo-transcriptome profiling of barley reveals induction of chitin elicitor receptor kinase gene (HvCERK1) conferring resistance against Fusarium graminearum

We report plausible disease resistance mechanisms induced by barley resistant genotype CI89831 against Fusarium head blight (FHB) based on metabolo-transcriptomics approach. We identified HvCERK1 as a candidate gene for FHB resistance, which is functional in resistant genotype CI9831 but non-functional in susceptible cultivars H106-371 and Zhedar-2. For the first time, we were able to show a hierarchy of regulatory genes that regulated downstream biosynthetic genes that eventually produced resistance related metabolites that reinforce the cell walls to contain the pathogen progress in plant. The HvCERK1 can be used for replacing in susceptible commercial cultivars, if non-functional, based on genome editing. Fusarium head blight (FHB) management is a great challenge in barley and wheat production worldwide. Though barley genome sequence and advanced omics technologies are available, till date none of the resistance mechanisms has been clearly deciphered. Hence, this study was aimed at identifying candidate gene(s) and elucidating resistance mechanisms induced by barley resistant genotype CI9831 based on integrated metabolomics and transcriptomics approach. Following Fusarium graminearum infection, we identified accumulation of specific set of induced secondary metabolites, belonging to phenylpropanoid, hydroxycinnamic acid (HCAA) and jasmonic acid pathways, and their biosynthetic genes. In association with these, receptor kinases such as chitin elicitor receptor kinase (HvCERK1) and protein kinases such as MAP kinase 3 (HvMPK3) and MAPK substrate 1 (HvMKS1), and transcription factors such as HvERF1/5, HvNAC42, HvWRKY23 and HvWRKY70 were also found upregulated with high fold change. Polymorphism studies across three barley genotypes confirmed the presence of mutations in HvCERK1 gene in two susceptible genotypes, isolating this gene as a potential candidate for FHB resistance. Further, the silencing of functional HvCERK1 gene in the resistant genotype CI9831, followed by gene expression and metabolite analysis revealed its role as an elicitor recognition receptor that triggered downstream regulatory genes, which in turn, regulated downstream metabolic pathway genes to biosynthesize resistance related (RR) metabolites to contain the pathogen to spikelet infection. A putative model on metabolic pathway regulation is proposed.

Integrated Metabolo-Transcriptomics Reveals Fusarium Head Blight Candidate Resistance Genes in Wheat QTL-Fhb2.

BackgroundFusarium head blight (FHB) caused by Fusarium graminearum not only causes severe losses in yield, but also reduces quality of wheat grain by accumulating mycotoxins. Breeding for host plant resistance is considered as the best strategy to manage FHB. Resistance in wheat to FHB is quantitative in nature, involving cumulative effects of many genes governing resistance. The poor understanding of genetics and lack of precise phenotyping has hindered the development of FHB resistant cultivars. Though more than 100 QTLs imparting FHB resistance have been reported, none discovered the specific genes localized within the QTL region, nor the underlying mechanisms of resistance.FindingsIn our study recombinant inbred lines (RILs) carrying resistant (R-RIL) and susceptible (S-RIL) alleles of QTL-Fhb2 were subjected to metabolome and transcriptome profiling to discover the candidate genes. Metabolome profiling detected a higher abundance of metabolites belonging to phenylpropanoid, lignin, glycerophospholipid, flavonoid, fatty acid, and terpenoid biosynthetic pathways in R-RIL than in S-RIL. Transcriptome analysis revealed up-regulation of several receptor kinases, transcription factors, signaling, mycotoxin detoxification and resistance related genes. The dissection of QTL-Fhb2 using flanking marker sequences, integrating metabolomic and transcriptomic datasets, identified 4-Coumarate: CoA ligase (4CL), callose synthase (CS), basic Helix Loop Helix (bHLH041) transcription factor, glutathione S-transferase (GST), ABC transporter-4 (ABC4) and cinnamyl alcohol dehydrogenase (CAD) as putative resistance genes localized within the QTL-Fhb2 region.ConclusionSome of the identified genes within the QTL region are associated with structural resistance through cell wall reinforcement, reducing the spread of pathogen through rachis within a spike and few other genes that detoxify DON, the virulence factor, thus eventually reducing disease severity. In conclusion, we report that the wheat resistance QTL-Fhb2 is associated with high rachis resistance through additive resistance effects of genes, based on cell wall enforcement and detoxification of DON. Following further functional characterization and validation, these resistance genes can be used to replace the genes in susceptible commercial cultivars, if nonfunctional, based on genome editing to improve FHB resistance.