Marker assisted backcrossing for introgression of Fusarium wilt resistance gene into melon

Fusarium wilt, caused by Fusarium oxysporum f. sp. melonis is a common vascular wilt fungal disease in melon across the world. The resistance gene to race 1 of this causal agent, Fom-2, has been previously cloned and its sequence is available. The objective of this research was the introgression of Fom-2 from one resistant (Isabelle) genotype into two susceptible cultivars (Garmak and Tile-torogh) via marker assisted backcrossing. First, the leucine-rich repeats (LRR) domain of Fom-2 from resistant and susceptible genotypes was sequenced to develop functional markers. A length of 1274 bp of the 3′ end of this gene was isolated, cloned and sequenced. The difference between resistant and susceptible genotypes in this region was 28 nucleotide substitutions. Two allele specific primer pairs, Fom2-R409 and Fom2-S253, were designed based on nucleotide substitutions to amplify resistant and susceptible alleles, respectively. For introgression of the gene, donor (Isabelle) and recurrent (Garmak and Tile-torogh) parents were crossed. Resistant plants in BC₁F₁ and BC₂F₁ generations were first detected using artificial pathogen inoculation and later the plants were genotyped by functional markers to validate their resistance. The resistant plants were also selected phenotypically in each generation for background genome recovery, which conduced to high similarity of BC₃ generation with the recurrent parents. It was proved the developed markers are more precise and efficient than inoculation trial and could be used as confident tools for screening of resistant melon genotypes to Fusarium wilt.     

Inheritance of race 1.2 Fusarium wilt resistance in four melon cultivars

The resistance to Fusarium oxysporum f.sp. melonis (Fom) race 1.2 has been studied in melons, such as the Portuguese accession ‘BG-5384’ and in the Japanese ‘Shiro Uri Okayama’, ‘Kogane Nashi Makuwa’, and ‘C-211’, since a good characterization of the resistance is necessary before its introgression into commercial varieties. These four melon accessions showed a high level of resistance to races 0, 1, and 2 of Fom, indicating that the partial resistance to the race 1.2 previously detected may not have been race specific. To determine the mode of inheritance of the resistance to Fom race 1.2, the F₁, F₂, BCP_R, and BCP_S generations from the crosses between the four resistant accessions above and ‘Piel de Sapo’, a Fom race 1.2 susceptible melon, were developed. They were subsequently inoculated with two Fom isolates, one from the pathotype 1.2Y and the other from the pathotype 1.2W. The area under the disease progress curve was determined for each inoculated plant, and the data were analyzed. We show that the resistance seen in these accessions is polygenically inherited with a complex genetic control because many epistatic interactions were detected. The three epistatic effects; additivity × additivity, dominance × dominance, and dominance × additivity are present and significant, with differing magnitudes from one cross to the next. The relatively low heritabilities, and these epistatic effects make difficult the improvement of the resistance, from these sources, through a standard selection procedure.     

Screening of Kabuli chickpea germplasm for resistance to Fusarium wilt

A total of 1915 Kabuli chickpea lines were screened in a wilt sick plot containing Fusarium oxysporum f.sp. ciceri race 0 at Béja, Tunisia. Complete resistance was found in 110 lines and this result was confirmed by a laboratory screening method. Principal components analysis showed that > 80% of the variation of the resistant lines was explained by hundred seed weight and days to maturity. Cluster analysis divided the resistant lines into four groups: 21 had high seed weight (48.25 ± 3.81 g) and early maturity (95.09 ± 2.50 d), 24 had high seed weight (46.84 ± 2.10 g) and late maturity (117.00 d), 34 had low seed weight (22.35 ± 4.72 g) and early maturity (92.97 ± 3.97 d) and 31 had low seed weight (19.62 ± 5.37 g) and late maturity (112.09 ± 4.51 d). This revised version was published online in July 2006 with corrections to the Cover Date.     

Breeding wheat and rye for resistance to Fusarium diseases

Fusarium culmorum and F. graminearum Groups 1 and 2 cause seedling blight, crown rot, foot rot and head blight in wheat and rye that may affect grain yield and quality for baking and feeding. This review starts with an analysis of Fusarium populations with regard to their genetic variation for aggressiveness, mycotoxin production, and isolate-by-host genotype interaction. To assess resistance in the different host growth stages, quantitative inoculation and disease assessment techniques are necessary. Based on estimated population parameters, breeding strategies are reviewed to improve Fusarium resistance in wheat and rye. Epidemiological and toxicological aspects of Fusarium resistance that are important for resistance breeding are discussed. F. culmorum and F. graminearum display large genetic variation for aggressiveness in isolate collections and in naturally occurring populations. The production of mycotoxins, especially deoxynivalenol and its derivatives, is a common trait in these populations. Significant isolate-by-host genotype interactions were not found across environments in wheat and rye. Artificial infections in the field are indispensable for improving Fusarium crown rot, foot rot and head blight resistance in wheat and rye. For a reliable disease assessment of large populations, disease severity ratings were found to be the most convenient. The differentiation of host resistance is greatly influenced by an array of nongenetic factors (macro-environment, microclimate, host growth stage, host organ) that show significant interactions with host genotype. Selection for environmentally stable resistance has to be performed in several environments under a maximum array of different infection levels. Selection in early growth stages or on one plant organ does not in most cases allow prediction of resistance in adult-plant stages or another plant organ. Significant genetic variation for resistance exists for all Fusarium-incited diseases in breeding populations of wheat and rye. The patho-systems studied displayed a prevalence of additive gene action with no consistent specific combining ability effects and thus rapid progress can be expected from recurrent selection. In wheat, intensive testing of parental genotypes allows good prediction of the mean head blight resistance after crossing. Subsequent selection during selfing generations enables the use of transgression towards resistance. In hybrid breeding of winter rye, the close correlation between foot rot resistance of inbred lines and their GCA effects implies that selection based on the lines per se should be highly effective. This is not valid for F. culmorum head blight of winter rye caused by a greater susceptibility of the inbred lines compared to their crosses. For both foot rot and head blight resistance, a high correlation between the resistance to F. graminearum and F. culmorum was found in wheat and rye. Mycotoxin accumulation occurs to a great extent in naturally and artificially infected plant stands. The correlation between resistance traits and mycotoxin contents are medium and highly dependent on the environment. Further experiments are needed to clarify whether greater resistance will lead to a correlated reduction of the mycotoxin content of the grains under natural infection.     

Studies on Cotton Breeding Resistant to Fusarium and Verticillium wilt Diseases

Both Fusarium and Verticillium wilts are important soil-borne diseases,which can not be effectively controlled by chemical fungicides.The two diseases,especially Verticillium wilt,have spread all over the cotton belt,and obstructed the progress of cotton production in China in recent years.It has been proven that breeding and growing resistant cultivars is one of the most economical and effective measures to control these diseases.The program of breeding cotton for resistance to wilt diseases has been continuously studied in Industrial Crops Research Institute,Sighuan Academy of Agricultural Sciences(ICRI-SAAS) for more than 50 years.     

Inheritance of resistance to Fusarium wilt (race 2) in chickpea

The inheritance of resistance to Fusarium wilt (race 2) of chickpea was studied in a set of three crosses, i.e. ‘WR315’בC104’ (resistant × susceptible), ‘WR315’בK850’ (resistant × tolerant) and ‘K850’בGW5/7’ (tolerant × tolerant) in order to investigate the number of genes involved, their complementation and to find out whether resistant segregants are possible in a cross between two tolerant cultivars. Tests of F₁, F₂ and F₃ generations of these crosses under controlled conditions at ICRISAT, Patancheru, India, indicated involvement of three loci (two recessive and one dominant alleles). The homozygous recessive form at the first two loci conferred resistance whereas susceptibility occurred when the first two loci were in the dominant form. A dominant allele at the third locus can complement the dominant alleles at the other two loci to confer tolerance. Occurrence of resistant segregants in a cross between two tolerant cultivars was observed.     

Fusarium vascular wilt in lentil: Inheritance and identification of DNA markers for resistance

The inheritance of resistance to lentil (Lens culinaris Medik.) vascular wilt caused by Fusarium oxysporum f.sp. lentis was investigated in a cross between resistant (ILL5588) and susceptible (L692–16-l(s)) lines. F_2:4 progenies and F_6:8, F_6:9 recombinant inbred line (RIL) populations were assessed for their wilt reaction for three seasons in a well-established wilt-sick plot. Resistance to wilt was conditioned by a single dominant gene in the populations studied. The map location of the Fw locus was identified for the first time through linkage to a random amplified polymorphic DNA (RAPD) marker (OPK-15₉₀₀) at 10.8 cM. Two other RAPD markers (OP-BH₈₀₀ and OP-DI5₅₀₀) identified by bulked segregant analysis were associated in the coupling phase with the resistance trait, and another marker (OP-C04₆₅₀) was associated with repulsion. The DNA markers reported here will provide a starting point in marker-assisted selection for vascular wilt resistance in lentil.     

Fusarium wilt of chickpea: physiological specialization, genetics of resistance and resistance gene tagging

Chickpea wilt caused by Fusarium oxysporum f. sp. ciceris is one of the major yield limiting factors in chickpea. The disease causes 10–90% yield losses annually in chickpea. Eight physiological races of the pathogen (0, 1A, 1B/C, 2, 3, 4, 5 and 6) are reported so far whereas additional races are suspected from India. The distribution pattern of these races in different parts of the world indicates regional specificity for their occurrence leading to the perception that F. oxysporum f. sp. ciceris evolved independently in different regions. Pathogen isolates also exhibit differences in disease symptoms. Races 0 and 1B/C cause yellowing syndrome whereas 1A, 2, 3, 4, 5 and 6 lead to wilting syndrome. Genetics of resistance to two races (1B/C and 6) is yet to be determined, however, for other races resistance is governed either by monogenes or oligogenes. The individual genes of oligogenic resistance mechanism delay onset of disease symptoms, a phenomenon called as late wilting. Slow wilting, i.e., slow development of disease after onset of disease symptoms also occurs in reaction to pathogen; however, its genetics are not known. Mapping of wilt resistance genes in chickpea is difficult because of minimal polymorphism; however, it has been facilitated to great extent by the development of sequence tagged microsatellite site (STMS) markers that have revealed significant interspecific and intraspecific polymorphism. Markers linked to six genes governing resistance to six races (0, 1A, 2, 3, 4 and 5) of the pathogen have been identified and their position on chickpea linkage maps elucidated. These genes lie in two separate clusters on two different chickpea linkage groups. While the gene for resistance to race 0 is situated on LG 5 of Winter et al. (Theoretical and Applied Genetics 101:1155–1163, 2000) those governing resistance to races 1A, 2, 3, 4 and 5 spanned a region of 8.2 cM on LG 2. The cluster of five resistance genes was further subdivided into two sub clusters of 2.8 cM and 2.0 cM, respectively. Map-based cloning can be used to isolate the six genes mapped so far; however, the region containing these genes needs additional markers to facilitate their isolation. Cloning of wilt resistance genes is desirable to study their evolution, mechanisms of resistance and their exploitation in wilt resistance breeding and wilt management.     

Testing strawberries for resistance to verticillium wilt

Summary The resistance to Verticillium wilt was investigated by inoculating the roots of runner plants and young seedlings of strawberry (Fragaria x ananassa Duch). In the glasshouse tests a satisfactory method was to raise the plants in jiffy pots and to remove the bottom of the pots prior to inoculation. This has the advantage that only slight root damage and no growth stagnation are caused.Both in the glasshouse and field tests inoculation resulted in a distinct infection reflected in reduced plant vigour and lesions on the petioles followed by wilting. In this way reliable differences between varieties could be demonstrated.The varieties grown in the glasshouse did not always exhibit the same degree of susceptibility as observed in practice. For example, Talisman and Redgauntlet were susceptible, while Gorella seemed to be more resistant because clear disease symptoms were not visible; the reduced plant vigour, however, indicated definite susceptibility. Generally the degree of infection and the reduction in growth were found to be significantly correlated.The two field trials carried out with plants the roots of which were infected by immersion in a suspension before planting, produced results which agreed better with practical findings than the glasshouse tests. In these trials Siletz appeared to be the most resistant variety, some American varieties and selections were nearly resistant, Vola and Juspa being not very susceptible.     

Variation in the response of melon genotypes to sudden wilt

Summary Differences in the response of melon genotypes to the sudden wilt disease were observed in several field trials conducted during 1993–1994 in the Arava region of southern Israel. Generally, the disease was more severe in the late summer growing season which is shorter and has higher temperatures than the spring and autumn growing seasons. The Oriental pickling melon breeding line P6a was the most tolerant among the entries tested. The response to the disease was also studied using two segregating families and their progenitors. BSK (tolerant) × P202 (susceptible) and P6a (tolerant) × D17 (susceptible). Wilting percentages of F₁, F₂ and backcross families were intermediate between the parents, suggesting an additive mode of gene action.     

Inheritance of resistance to Fusarium wilt in Indian lentil germplasm (Lens culinaris Medik.)

Summary Three lentil genotypes resistant to Fusarium oxysporum f.sp. lentis viz. Pant L 234, JL 446 and LP 286 were crossed with two susceptible ones. The hybrid plants were all resistant in the eight crosses evaluated. Segregation pattern for wilt reaction in F₂, BC(P₁), BC(P₂) and F₃ generations in field and glasshouse conditions indicated that resistance to Fusarium wilt is under the control of two dominant duplicate genes in Pant L 234 and two independent dominant genes with complementary effects in JL 446 and LP 286. A third dominant gene complementary to the dominant genes in JL 446 and LP 286 is present in two susceptible lines. Allelic tests suggest the presence of five independently segregating genes for resistance. Duplicate dominant genes in Pant L 234 are non-allelic to two dominant genes with complementary effects in LP 286 and JL 446 and the third gene complementary to the two genes in JL 446 and LP 286 in susceptible lines JL 641 and L 9–12. Gene symbols among parental genotypes have been designated.     

Screening lentil for resistance to fusarium wilt: methodology and sources of resistance

Host-plant resistance is the best means to control the key disease of lentil-vascular wilt, caused by Fusarium oxysporum Schlecht. emend. Snyder & Hansen f.sp. lentis Vasudeva and Srinivasan. Systematic screening for resistance to lentil wilt was initiated in the field in 1993, in a wilt-sick plot in North Syria, with a core collection of 577 germplasm accessions from 33 countries. A subset (88 accessions) of mostly resistant accessions was re-screened in 1994. The most resistant accessions came from Chile, Egypt, India, Iran and Romania. Variation among accessions in the temporal pattern of wilting was analyzed. The limited wilting in resistant accessions followed a linear model through time, whereas the pattern for susceptible accessions was better described with an exponential model. This temporal variation emphasizes the need for repeated scoring during screening for resistance to lentil vascular wilt to identify ‘late-wilters’. To overcome spatial variation in inoculum density, the efficacy of using wilt scores from a systematically-repeated susceptible control in covariate analysis was tested. Covariance analysis significantly improved overall screening by 3% in 1993, but the improvement was non-significant in 1994. The results emphasize the relative uniformity of disease pressure in the wilt-sick plot and suggest that covariance analysis of a systematically arranged control will be of greater benefit in land which is less uniformly infected. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification of dominant and recessive genes for resistance to Fusarium wilt in pigeonpea and their implication in breeding hybrids

Fusarium wilt is an important disease of pigeonpea [Cajanus cajan (L.) Millsp.] and it can cause severe yield losses. Chemical control of this disease is difficult and expensive; therefore, cultivation of resistant varieties/hybrids is the most efficient strategy for enhancing the production. In the present study, by using a wilt susceptible cytoplasmic-nuclear male-sterile line and four wilt resistant fertility restorers, one dominant and one recessive gene with dominant suppressive epistatic effects were found responsible for controlling resistance to Fusarium wilt. Considering the annual losses and wide spread nature of wilt diseases in pigeonpea, it is imperative that all the inbred and hybrid cultivars have high level of resistance to this disease. The presence of dominant gene for resistance will increase the efficiency of breeding wilt resistant cultivars because it will yield greater proportion of resistant genotypes in segregating generations. In hybrid breeding also, the presence of dominant gene for wilt resistance will be an advantage. The transfer of this gene in female hybrid parents will ease the breeding of wilt resistant hybrids because this will allow the use of both wilt resistant as well as susceptible restorers in generating wilt resistant hybrid combinations.     

A second gene for resistance to race 4 of Fusarium wilt in chickpea and linkage with a RAPD marker

Fusarium wilt caused by Fusarium oxysporum Schlechtend.: Fr f. sp. ciceris (Padwick) Matuo & Sato is a devastating disease of chickpea. The current study was conducted to determine the inheritance of the gene(s) for resistance to race 4 of fusarium wilt and to identify linked RAPD markers using an early wilting line, JG-62, as a susceptible parent. Genetic analysis was performed on the F₁s, F₂s and F₃ families from the cross of JG-62 × Surutato-77. The F₃ families were inoculated with a spore suspension of the race 4 wilt pathogen and the results were used to infer the genotypes of the parent F₂ plants. Results indicated that two independent genes controlled resistance to race 4. Linkage analysis of candidate RAPD marker, CS-27₇₀₀, and the inferred F₂ phenotypic data showed that this marker locus is linked to one of the resistance genes. Allelism indicated that the two resistance sources, Surutato-77 and WR-315, shared common alleles for resistance and the two susceptible genotypes, C-104 and JG-62, carried alleles for susceptibility. The PCR-based marker, CS-27₇₀₀, was previously reported to be linked to the gene for resistance to race 1 in a different population which suggested that the genes for resistance to races 1 and 4 are in close proximity in the Cicer genome. This revised version was published online in August 2006 with corrections to the Cover Date.     

Breeding and genetics of Fusarium basal rot resistance in onion

Fusarium basal plate rot (FBR), caused by Fusarium oxysporum f. sp. cepae, is an important soil-borne disease of onions worldwide. The causal organism infects the basal stem plate of the bulb and eventually kills the entire plant through degradation of the basal plate. F. o. f. sp.cepae infections in dormant bulbs during storage allow secondary infections to occur. The primary method of infection by F. o. f. sp. cepaeis through direct penetration of the basal stem plate. Infection can also occur through wounded tissue particularly roots and basal portions of bulb scales. The most cost-effective methods of control are crop rotation and host plant resistance. Current research suggests that a single gene, two genes, or multiple genes govern resistance to FBR. Breeding programs have successfully used screening procedures to develop intermediate- and long-day, FBR-resistant cultivars. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic analysis of Fusarium wilt resistance in cowpea (Vigna unguiculata Walp.)

This study investigated the inheritance of resistance to Fusarium oxysporum f.sp. tracheiphilum (Fot) in cowpea lines. Resistant and susceptible cowpea lines were crossed to develop F₁, F₂ and backcross populations. Reaction to Fot was evaluated in 2015 and 2016 using seed soak and modified root‐dip inoculation methods. The expression of resistance reaction in the F₁ and segregation in F₂ generations indicated the role of dominant gene controlling Fot in cowpea. These results were further supported by the result of backcross (BC₁P₁F₁ and BC₁P₂F₁) progeny tests. The backcross of F₁ with the resistant parent produced progeny that were uniformly resistant, whereas backcross of F₁ with the susceptible parent produced progeny that segregated into 1:1 ratio. The F₂ segregation ratio in the reciprocal cross showed no evidence of maternal effect in the inheritance of the resistance. Allelism test suggests that the gene for resistance in TVu 134 was the same in TVu 410 and TVu 109‐1. We also identified an SSR marker, C13‐16, that cosegregated with the gene conferring resistance to Fot in cowpea.     

Assessment of Gossypium sturtianum and G. australe as potential sources of Fusarium wilt resistance to cotton

When challenged with Fusarium oxysporum f. sp. vasinfectum (Fov) from vegetative compatibility groups (VCGs) 01111 and 01112 in glasshouse tests, Gossypium australe Mueller and Gossypium sturtianum Willis accessions showed a variety of disease responses ranging from highly resistant to highly susceptible. Under high disease pressure G. sturtianum accession Gos-5275 was significantly more resistant than the commercial G. hirsutum cultivars that are designated standards for Fusarium resistance by Australian cotton breeders. Under low disease pressure G. sturtianum accession Gos-5250 was more susceptible than a highly susceptible commercial cultivar. A series of glasshouse tests was performed at two locations (Indooroopilly, QLD. and Canberra, ACT), and under low and high disease pressure. In these tests, a hexaploid cross (Gos-5271) generated from a Fusarium-resistant G. sturtianum (Gos-5275) and a Fusarium-susceptible G. hirsutum L. (CPI-138969) was significantly more resistant to Fusarium wilt than its G. hirsutum parent. Thus G. sturtianum, with a diploid genome and a range of responses to Fov challenge, has the potential to provide the basis for the elucidation of the genetic basis of resistance to Fusarium wilt in cotton species. In addition, resistant accessions of G. sturtianum are identified as a potential source of Fusarium wilt resistance genes for cotton breeding. In the glasshouse tests used to assess the resistance of various Gossypium accessions to Fusarium wilt disease, the scoring of vascular browning was found to give a more reliable indication of disease severity than the scoring of foliar symptoms. This revised version was published online in August 2006 with corrections to the Cover Date.     

Breeding maize for resistance to streak

A historical review is given of the breeding for resistance to streak disease in Maize. The latest advances in this respect in South Africa include the production of a commercial streak tolerant three-way hybrid, S.A. 31, in which the resistance to streak is derived from P x H maize. New sources of resistance have been found in Mexican maize, the inbreds Urg. 54 and Mex. 37-5 being the most promising.

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Samenvatting Een historisch overzicht wordt gegeven van het kweken op resistentie tegen de door een virus veroorzaakte strepenziekte (streak-disease) bij maïs.Het gelukte in Zuid-Afrika de tolerante driestam-hybride SA 31 te kweken, die de resistentie tegen strepenziekte dankt aan de P×H hybride.Nieuwe bronnen van resistentie zijn gevonden in Mexicaanse maïs, waarvan de inteeltstammen Urg.54 en Mex.37-5 veelbelovend zijn.