Genetic mapping of bph20(t) and bph21(t) loci conferring brown planthopper resistance to Nilaparvata lugens St?l in rice (Oryza sativa L.)

Brown planthopper(BPH) is one of the most serious and destructive insect pests of rice in most rice growing regions of the world. In this study, two major resistance genes against BPH have been identified in an Oryza rufipogon (Griff.) introgression rice line, RBPH54. Inheritance of the BPH resistance in RBPH54 was studied by screening the resistance in parents, F1, F2 and BC1 generations against BPH biotype 2. A population of BC3F2 lines was developed and SSR markers were employed for the gene mapping, and new markers were designed for fine mapping of the resistance genes, while sequence information of BAC/PAC clones was used to construct physical maps of the genes. The results showed that the BPH resistance in RBPH54 was governed by recessive alleles at two loci, tentatively designated as bph20(t) and bph21(t). The locus bph20(t) was fine mapped to the short arm of chromosome 6 about 2.7 cM to the upper marker RM435 and 2.5 cM to lower marker RM540 and in a 2.5 cM region flanked by two new SSR markers BYL7 and BYL8 which were developed in the present study. The other BPH resistance locus bph21(t) was initially mapped to a region 7.9 cM to upper marker RM222 and 4.0 cM to lower marker RM244 on the short arm of chromosome 10. For physical mapping, the bph20(t)-linked markers were landed on BAC/PAC clones of the reference cv., Nipponbare, released by the International Rice Genome Sequencing Project. The bph20(t) locus was physically defined to an interval of about 75 kb with clone P0514G1. Identification and location of these two genes in the present study have diversified the BPH resistance gene pool, which give benefit to the development of resistant rice cultivars, and the linkage PCR-based SSR markers for the bph20(t) and bph21(t) genes would help realize the application of the genes in rice breeding through marker-assisted selection.     

Genetic basis of multiple resistance to the brown planthopper (Nilaparvata lugens St?l) and the green rice leafhopper (Nephotettix cincticeps Uhler) in the rice cultivar ‘ASD7’ (Oryza sativa L. ssp. indica)

The rice cultivar ASD7 (Oryza sativa L. ssp. indica) is resistant to the brown planthopper (BPH; Nilaparvata lugens Stål) and the green leafhopper (Nephotettix virescens Distant). Here, we analyzed multiple genetic resistance to BPH and the green rice leafhopper (GRH; Nephotettix cincticeps Uhler). Using two independent F₂ populations derived from a cross between ASD7 and Taichung 65 (Oryza sativa ssp. japonica), we detected two QTLs (qBPH6 and qBPH12) for resistance to BPH and one QTL (qGRH5) for resistance to GRH. Linkage analysis in BC₂F₃ populations revealed that qBPH12 controlled resistance to BPH and co-segregated with SSR markers RM28466 and RM7376 in plants homozygous for the ASD7 allele at qBPH6. Plants homozygous for the ASD7 alleles at both QTLs showed a much faster antibiosis response to BPH than plants homozygous at only one of these QTLs. It revealed that epistatic interaction between qBPH6 and qBPH12 is the basis of resistance to BPH in ASD7. In addition, qGRH5 controlled resistance to GRH and co-segregated with SSR markers RM6082 and RM3381. qGRH5 is identical to GRH1. Thus, we clarified the genetic basis of multiple resistance of ASD7 to BPH and GRH.     

Close linkage of a blast resistance gene,Pias(t), with a bacterial leaf blight resistance gene,Xa1-as(t), in a rice cultivar ‘Asominori’

It has long been known that a bacterial leaf blight-resistant line in rice obtained from a crossing using ‘Asominori’ as a resistant parent also has resistance to blast, but a blast resistance gene in ‘Asominori’ has not been investigated in detail. In the present study, a blast resistance gene in ‘Asominori’, tentatively named Pias(t), was revealed to be located within 162-kb region between DNA markers YX4-3 and NX4-1 on chromosome 4 and to be linked with an ‘Asominori’ allele of the bacterial leaf blight resistance gene Xa1, tentatively named Xa1-as(t). An ‘Asominori’ allele of Pias(t) was found to be dominant and difference of disease severity between lines having the ‘Asominori’ allele of Pias(t) and those without it was 1.2 in disease index from 0 to 10. Pias(t) was also closely linked with the Ph gene controlling phenol reaction, suggesting the possibility of successful selection of blast resistance using the phenol reaction. Since blast-resistant commercial cultivars have been developed using ‘Asominori’ as a parent, Pias(t) is considered to be a useful gene in rice breeding for blast resistance.     

Identification of genomic regions linked to blast (Pyricularia grisea) resistance in pearl millet

Blast disease causes serious economic yield losses in pearl millet. Identification and introgression of genomic regions associated with blast resistance can help to develop resistant cultivars to minimize yield losses incurred from blast outbreaks. In this study, 384 advanced pearl millet genotypes were screened against six blast pathotype-isolates (major pearl millet growing agro-ecologies of India), namely, Pg 45, Pg 118, Pg 138, Pg 186, Pg 204 and Pg 232. Analysis of variance showed significant (P < .001) variation among genotypes for blast reaction (susceptible to resistance). ICMR 08111 and ICMR 10888 genotypes showed resistance to all six blast pathotypes. A genome-wide association study performed with 264,241 single nucleotide polymorphic markers could successfully identify 15 SNPs (P = 1.26 × 10⁻⁷ to 9.22 × 10⁻¹²) underlying the genomic regions governing blast-resistance across five different chromosomes. The SNPs reported had a significant association in at least two of the three models tested (GLM, MLM and Farm CPU). These SNPs can be used in pearl millet-resistant breeding programmes after their function has been validated across different genetic backgrounds.     

Dynamic controlled atmosphere (DCA): Does fluorescence reflect physiology in storage?

A link between the minimum fluorescence (F_o) and a metabolic shift from predominantly aerobic to fermentative metabolism [i.e. the lower oxygen limit (LOL)] is the foundation of dynamic controlled atmosphere (DCA). Current DCA technology uses pulse frequency modulated (PFM) sensors and employs a range of light intensities and extrapolation to measure F_α, an approximation of F_o. Like fruit mass, colour, sugar or acid levels, the LOL is inherently variable, even between apples (Malus domestica) (for example) from a given cultivar and tree or between the sun-exposed and shaded regions of a single fruit. The physiological link between metabolism and fluorescence has not been extensively studied. However, recent work suggests the low-O₂-induced rise in F_α results from a shut down of mitochondrial function and a buildup of reductant that leads to an over-reduction of the plastoquinone (PQ) pool and a decrease in photochemical quenching. Hypoxic conditions above the LOL can decrease F_α slightly in some species, possibly as a result of zeaxanthin formation and increased non-photochemical quenching. Low-intensity light differentially affects F_α depending on the O₂ level: light increases F_α when O₂ levels are above the LOL due to light-induced reduction of the oxidized PQ pool, but decreases the elevated F_α signal below the LOL as a result of a PSI-driven oxidation of the over-reduced PQ pool. Temperature has a negative, primarily non-physiological correlation with the F_α baseline which seems unrelated to the PQ pool redox state. Understanding how O₂ and other factors affect F_α may improve the utility and commercial application of DCA.     

A new gene <Emphasis Type="Italic">Bph33</Emphasis>(<Emphasis Type="Italic">t</Emphasis>) conferring resistance to brown planthopper (BPH), <Emphasis Type="Italic">Nilaparvata lugens</Emphasis> (Stål) in rice line RP2068-18-3-5

The rice brown planthopper (BPH) Nilaparvata lugens (Stål) is one of the major pests of rice across Asia. Host-plant resistance is the most ecologically acceptable means to manage this pest. A rice breeding line RP2068-18-3-5 (RP2068) derived from the land race Velluthacheera is reported to be resistant to BPH populations across India. We identified a new R gene [Bph33(t)] in this line using advanced generation RILs derived from TN1 × RP2068 cross through phenotyping at two locations and linkage analysis with 99 polymorphic SSR markers. QTL analysis through IciMapping identified at least two major QTL on chromosome 1 influencing seedling damage score in seed box screening, honey dew excretion by adults and nymphal survival. Since no BPH R gene has been reported on chromosome 1, we designate this locus as a new gene Bph33(t) which accounted for over 20% of phenotypic variance. Scanning the region for candidate gene suggested two likely candidates a leucine rich repeat (LRR) gene and a heat shock protein (HSP) coding gene. Expression profiling of the two genes in the two contrasting parents and RILs showed induction of the HSP gene (LOC_Os01g42190.1) at 6 h after infestation while LRR gene did not show such induction. It is likely that the HSP represented Bph33(t).     

Delineating Genotype × Environment interactions towards durable resistance in mungbean against Cercospora leaf spot (Cercospora canescens) using GGE biplot

Cercospora leaf spot (Cercospora canescens) is a major fungal disease which impedes mungbean production worldwide. Presence of wider host range with existence of pathogenic variability creates intricacy towards host-pathogen dynamics. Moreover, environmental factors having crucial role in augmenting severity of this disease further complicate disease management. An attempt has been made for unfolding genotype x environment interactions towards identifying and validating durable resistant genotypes against cercospora leaf spot in multi-environment testing. Preliminary screening with 246 genotypes under artificial epiphytotic condition was conducted to extract out a subset of 22 mungbean genotypes for further evaluation in field testing across six environments consecutively for two years. GGE biplot analysis detected significant environmental influence towards genotypic response and confirmed the presence of non-crossover interaction with incoherent genotypic response, thus advocating the urgency for multi-location testing. GGE biplot aptly identified “LGG 460” and “COGG 912” as “ideal” and “desirable” genotypes, respectively having durable resistance and genetic homeostasis and thus suggested for their utilization in future resistance breeding programme in mungbean against cercospora leaf spot.     

Development of bio-assays and screening for resistance to beet armyworm (Spodoptera Exigua Hübner) in Allium cepa L. and its wild relatives

The beet armyworm (Spodoptera exigua Hübner)is the most important pest in tropical Alliumcultivations. All shallot (Allium cepa L. group Aggregatum) cultivars are susceptible to this pest. Therefore accessions from three wild Alliumspecies, namely A. galanthum Kar. et Kir., A. fistulosum L. and A. royleiStearn, next to A. cepa L. were used to screen for resistance. First of all, a reliable bio-assay had to be developed. To this end transparent plastic cages with in total 5 plants of one accession per cage were placed on per lite in a heated greenhouse. Five 3-day old larvae were inoculated on each plant. Eight days after inoculation the number of surviving larvae per cage and the mean fresh weightper larva was determined. The lowest larval survival (36%) was found on A. roylei. This was not, however, significantly different from other Allium accessions. Significant differences were found in the fresh weight per larva fed on different Allium accessions. The larvae survived on A. roylei had a very low fresh weight (10.3 mg per larva), while those on an accession of A. fistulosum had the highest fresh weight (45.1 mg per larva). The larval fresh weight on A. roylei was lower than all the other accessions except from the tropical shallot cultivar Bawang Bali. To check whether or not a toxic compound was involved in the resistance present in A. roylei, tenaccessions from four Allium species were screened. Five 3-day old larvae were inoculated on regularly replaced leaf material of each accession ofAllium species. No significant differences were found in mean fresh weight per larva and mean survival of larvae among different accessions. There were also no significant differences in pupal weight and developmental time. All larvae became pupae 10 days after inoculation. The data indicate that there is no toxic compound present in A. roylei. These results are underlined by the observation in the greenhouse bio-assay that A. roylei plants were equally damaged by the beet armyworm compared to otherAllium species. The results obtained so far therefore suggest that introduction of resistance to S. exigua via the exploitation of variation for resistance to the beet armyworm in A. roylei is unclear and that genetic engineering using Cry sequences could provide a way forward. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Quantitative trait locus mapping for Verticillium wilt resistance in a backcross inbred line population of cotton (Gossypium hirsutum × Gossypium barbadense) based on RGA-AFLP analysis

Verticillium wilt (VW), caused by Verticillium dahliae Kleb., is one of the most important diseases in cotton. The objective of this study was to map quantitative trait loci (QTLs) conferring VW resistance using resistance gene analog (RGA)-targeted amplified fragment length polymorphism (RGA-AFLP) markers in an interspecific backcross inbred line mapping population, consisting of 146 lines from a susceptible Sure-Grow 747 (Gossypium hirsutum L.) × resistant Pima S-7 (G. barbadense L.) cross. VW resistance was evaluated in replicated tests based on disease incidence in the field, and disease incidence and severity in the greenhouse. Of 160 polymorphic RGA-AFLP markers, 42 were significantly correlated with one or more VW traits and 41 were placed on a linkage map which covered 1,226 cM of the cotton genome and contained 251 other molecular markers. Three QTLs for VW resistance were detected, each of which explained 12.0–18.6 % of the phenotypic variation. Two of these QTLs for disease incidence and severity detected in the greenhouse inoculation tests using root wounding are located on chromosome c4. Both are closely linked to four RGA-AFLP markers and therefore considered as the same QTL for VW resistance. The other QTL detected in the field test was located on c19 and flanked by several RGA-AFLP markers. The desirable QTL allele on c4 for VW resistance detected in the greenhouse was from the VW susceptible Upland parent and absent from the resistant Pima parent which was more VW susceptible due to the disarmament of the first line of defense mechanism due to root wounding during inoculation. The other desirable VW resistance QTL allele, on c19, was from the resistant parent Pima S-7, consistent with the fact that Pima cotton was more resistant to VW when naturally infected in the field. The results should facilitate the development of more sequence specific markers and the transfer of VW resistance from Pima to Upland cotton through marker-assisted selection.     

Great Recession and club convergence in Europe: A cross-country,cross-region panel analysis (2000–2015)

The paper aims at investigating the impact of the Great Recession on per capita GDP convergence process across European regions and countries. Using the time-varying factor model developed by Phillips and Sul for the period 2000–2015 and two different merging procedures to identify clubs, we provide evidence of the diverging impact of the Great Recession “between” the higher and the lower convergence clubs at both regional and country levels as well as of the strengthening of the convergence process “within” most clubs. In addition, we add further evidence to the common belief of a “multi-speed” Europe by contrasting Eastern European countries' and regions' behavior vis-à-vis original European members' one, and by identifying the factors that affect club membership and resilience to the recent economic downturn. We find that the membership in the higher clubs and resilience to the Great Recession are positively affected by the presence of several local-specific factors and macroeconomic characteristics.     

Genetic analysis and genotype × environment (G × E) for grey leaf spot disease resistance in elite African maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) germplasm

Maize grey leaf spot (GLS) disease remains an important foliar disease in sub-Saharan Africa accounting for more than 25% yield losses in maize. Information on inheritance of GLS resistance of germplasm adapted to African environments is required in new sources being identified. Therefore, hybrids generated from a 10 × 10 half-diallel mating of tropical advanced maize inbred lines were evaluated in six environments to determine combining ability, genotype × environment interaction (G × E) and the impact of GLS disease on grain yield. General combining ability effects were highly significant and accounted for 72 and 68% of the variation for GLS resistance and grain yield, respectively. Significant specific combining ability effects associated with reduced disease levels were observed in some hybrids when one parent was resistant, and these may be exploited in developing single cross maize hybrids. Regression analysis showed a 260–320 kg ha^?1 decrease in maize grain yield per each increase in GLS disease severity score, and significant associations (r = ?0.31 to ?0.60) were observed between grain yield and GLS severity scores. This showed the potential of GLS disease to reduce yield in susceptible varieties grown under favourable disease conditions, without control measures. Genotype and genotype × environment biplots and correlation analysis indicated that the significant G × E observed was not due to changes in hybrid ranking, implying absence of a significant crossover interaction. Therefore, predominance of additive gene effects imply that breeding progress for GLS disease resistance would be made through selection and this could be achieved at a few hot-spot sites, such as Baynesfield and Cedara locations in South Africa, and still deploy the resistant germplasm to other environments in which they are adapted.     

Resistance to rice sheath blight (<Emphasis Type="Italic">Rhizoctonia solani</Emphasis> Kühn) [(teleomorph: <Emphasis Type="Italic">Thanatephorus cucumeris</Emphasis> (A.B. Frank) Donk.] disease: current status and perspectives

Sheath blight (ShB) disease, caused by Rhizoctonia solani, is an economically important rice disease worldwide, especially in intensive production systems. Several studies have been conducted to identify sources for ShB resistance in different species of rice, including local accessions and landraces. To date, none of the genotypes screened are immune to ShB, although variation in levels of resistance have been reported. Several quantitative trait loci (QTL) for ShB resistance have been identified using mapping populations derived from indica or japonica rice. A total of 33 QTL associated with ShB resistance located on all 12 rice chromosomes have been reported, with ten of these co-localizing with QTL for morphological attributes, especially plant height, or for heading date. Sixteen QTL, from the same or differing genetic backgrounds, have been mapped at least twice. Of these, nine QTL were independent of morphological traits and heading date. We hypothesize that two main, distinct, mechanisms contribute to ShB resistance: physiological resistance and disease escape. Strategies to improve our understanding of the genetics of resistance to ShB are discussed.     

Oral Inoculation with O157:H7(△hly△stx△toxB) Leads to Decreased Shedding in Goats after Challenge with Escherichia coli O157 :H7

为了明确O157:H7(△hly △stx△toxB)口服能否减少亲本株O157:H7在山羊体内的定殖.选用3月龄山羊,首先口服接种O157:H7(△hly△stx△toxB),分别于第7,14天再次口服接种亲本株O157:H7,监测亲本株O157:H7的排菌持续时间和排菌量.结果表明:O157:H7(△hly △stx△toxB)口服后第7天攻击亲本株O157:H7,第4天检测不到亲本株O157:H7的排菌;第14天攻击亲本株O157:H7,第9天检测不到亲本株O157:H7的排菌.直接口服亲本株O157:H7,排菌一直持续到第25天.口服O157:H7(△hly △stx△toxB)能够减少O157:H7在山羊体内的定殖,为研制EHEC O157:H7基因缺失疫苗奠定了基础.     

Identification of molecular markers associated with resistance to squash silverleaf disorder in summer squash (<Emphasis Type="Italic">Cucurbita</Emphasis> <Emphasis Type="Italic">pepo</Emphasis>)

Squash silverleaf (SSL), caused by the silverleaf whitefly [Bemisia argentifolii (formerly known as Bemisia tabaci Gennadius, B strain)], is an important physiological disorder that affects squash (Cucurbita spp.) by reducing yield potential. Breeding squash with resistance to SSL disorder can be facilitated by using marker-assisted selection (MAS). Resistance to SSL disorder, in Cucurbita pepo, is conferred by a single recessive gene (sl). The objective of this study was to identify molecular markers associated with resistance. A zucchini squash, SSL disorder resistant breeding line, ‘Zuc76’ (sl/sl) and a SSL disorder susceptible zucchini cultivar ‘Black Beauty’ (Sl/Sl) were screened with 1,152 randomly amplified polymorphic DNA (RAPD) primers and 432 simple sequence repeat (SSR) markers to identify polymorphisms. Using F₂ and BC₁ progeny segregating for SSL disorder resistance, three RAPD (OPC07, OPL07 and OPBC16) primers and one SSR (M121) marker were found associated with sl. Fragments amplified by RAPD primer OPC07 was linked in coupling phase to sl, whereas RAPD primer OPL07 was linked in repulsion phase. RAPD primer OPBC16 and SSR marker M121 were co-dominant. The allelic order of these loci was found to be M121–sl–OPC07–OPL07–OPBC16. The closest marker to sl is M121 with an estimated genetic distance of 3.3 cM. The markers identified in this study will be useful for breeding summer squash (C. pepo) for SSL disorder resistance derived from zucchini squash breeding line ‘Zuc76’.     

Inheritance of foliar resistance to <Emphasis Type="Italic">Phytophthora</Emphasis> species in cacao (<Emphasis Type="Italic">Theobroma cacao</Emphasis> L.): can it be influenced by CSSV infection?

Studies on quantitative genetics of foliar resistance to black pod disease in cacao could inadvertently use cocoa swollen shoot virus (CSSV) infected leaves which could bias the results especially in West Africa where the virus is prevalent. However, effects of CSSV on inheritance and heritability of foliar resistance to Phytophthora species is not known. Choice of an efficient breeding method requires an accurate estimation of genetic effects in selection schemes for foliar resistance to Phytophthora species in cacao. The objective of this study was to investigate the effect of CSSV infection on quantitative genetic parameters of foliar resistance to cocoa black pod disease in a population of 36 F1 hybrids developed by mating six cacao genotypes using a diallel method. The generated F1s and their parents were evaluated for foliar resistance to P. palmivora and P. megakarya using a randomized complete block design (RCBD) with three replications. 1A CSSV and Nsaba CSSV strains were used to infect the cacao genotypes using the patch graft method. The parents chosen showed significant variations for scores of leaf discs after inoculation with P. palmivora and P. megakarya. The leaf disc scores of CSSV infected crosses were lower than leaf disc scores of CSSV-free crosses. Genetic component analysis showed that the effects of GCA and SCA was significant for both CSSV-free and CSSV-infected crosses in resistance to P. palmivora and P. megakarya. The significant GCA and SCA for both CSSV-free and CSSV-infected crosses strongly suggest that both additive and non-additive genetic effects play an important role in the determination of inheritance of foliar resistance to Phytophthora species in cacao. There was significant variability in mean squares of GCA and SCA of CSSV-free and CSSV-infected crosses indicating that CSSV infection modifies GCA and SCA of affected plants. Narrow sense heritability was relatively low (0.31) for foliar resistance to P. palmivora and P. megakarya under CSSV-free and 1A CSSV strain infected conditions. However, heritability for foliar resistance to P. palmivora (0.43) and P. megakarya (0.36) was significantly higher under Nsaba CSSV infected condition. The modifications of mean squares of GCA and SCA and narrow sense heritability due to CSSV infection could mislead in choice of breeding methods indicating that attention must be paid to the infection status of plants when conducting quantitative genetics studies using diseased and healthy plants. CSSV status of leaf samples should be known before using them in leaf disc test. Genotypes Pa7/808 and Pound 7 had high negative GCA effects and are promising parents for enhancement of resistance to black pod disease in cacao.     

Estimation of outcrossing rates in intraspecific (<Emphasis Type="Italic">Oryza sativa</Emphasis>) and interspecific (<Emphasis Type="Italic">Oryza sativa</Emphasis> × <Emphasis Type="Italic">Oryza glaberrima</Emphasis>) rice under field conditions using agro-morphological markers

Rice is mainly a self-pollinating crop, but some outcrossing has been reported. Outcrossing with an undesirable donor would lead to the creation of segregants or off-types, which would adversely affect genetic purity and uniformity of the crop. Outcrossing rates in rice under field conditions were investigated using cultivar WAB96-1-1 as a pollen donor and WAB56-104, NERICA 2, NERICA 4 and NERICA 7 as pollen recipients. Levels of outcrossing were investigated up to 30 m from the pollen donor. Dominant morphological markers of red kernel colour and pubescent leaves of the donor were used to identify hybrids. A total of 721 134 plants were investigated. There was an average outcrossing rate of 0.7 ± 0.51%, with a potential outcrossing rate of 2.45 ± 0.86%. Outcrossing rates decreased with increase in distance. It ranged from 2.45% at 0.2 m from the donor to 0.05% at 25 m from the donor. Differences were observed between genotypes and seasons. In season 1 the highest average outcrossing rate of 1.2 ± 0.63% was with WAB56-104 and in season 2 it was 1.1 ± 0.69% with NERICA 4. Outcrossing occurred up to 30 m from the donor. This has implications for germplasm management and conservation and the production of high quality seed. Spatial isolation remains the most practical method to prevent undesirable gene flow. The study indicated that red kernel colour and leaf pubescence can be used to effectively assess outcrossing under field conditions in rice.