Identification and mapping of H32, a new wheat gene conferring resistance to Hessian fly

H32 is a newly identified gene that confers resistance to the highly pervasive Biotype L of the Hessian fly [ Mayetiola destructor (Say)]. The gene was identified in a synthetic amphihexaploid wheat, W-7984, that was constructed from the durum ‘Altar 84’ and Aegilops tauschii. This synthetic wheat is one of the parents of the marker-rich ITMI population, which consists of 150 recombinant inbred lines (RILs) derived by single-seed descent from a cross with ‘Opata 85’. Linkage analysis of the H32 locus in the ITMI population placed the gene between flanking microsatellite (SSR) markers, Xgwm3 and Xcfd223, at distances of 3.7 and 1.7 cM, respectively, on the long arm of chromosome 3D. The Xgwm3 primers amplified codominant SSR alleles, a 72 bp fragment linked in coupling to the resistance allele and an 84 bp fragment linked in repulsion. Primers for the SSR Xcfd223 amplified a 153 bp fragment from the resistant Synthetic parent and a 183 bp fragment from the susceptible Opata line. Deletion mapping of the flanking Xgwm3 and Xcfd223 markers located them within the 3DL-3 deletion on the distal 19% of the long arm of chromosome 3D. This location is at least 20 cM proximal to the reported 3DL location of H24, a gene that confers resistance to Biotype D of the Hessian fly. Tight linkage of the markers will provide a means of detecting H32 presence in marker-assisted selection and gene pyramiding as an effective strategy for extending durability of deployed resistance.     

Precisely mapping a major gene conferring resistance to Hessian fly in bread wheat using genotyping-by-sequencing

Background

One of the reasons hard red winter wheat cultivar ‘Duster’ (PI 644016) is widely grown in the southern Great Plains is that it confers a consistently high level of resistance to biotype GP of Hessian fly (Hf). However, little is known about the genetic mechanism underlying Hf resistance in Duster. This study aimed to unravel complex structures of the Hf region on chromosome 1AS in wheat by using genotyping-by-sequencing (GBS) markers and single nucleotide polymorphism (SNP) markers.

Results

Doubled haploid (DH) lines generated from a cross between two winter wheat cultivars, ‘Duster’ and ‘Billings’ , were used to identify genes in Duster responsible for effective and consistent resistance to Hf. Segregation in reaction of the 282 DH lines to Hf biotype GP fit a one-gene model. The DH population was genotyped using 2,358 markers developed using the GBS approach. A major QTL, explaining 88% of the total phenotypic variation, was mapped to a chromosome region that spanned 178 cM and contained 205 GBS markers plus 1 SSR marker and 1 gene marker, with 0.86 cM per marker in genetic distance. The analyses of GBS marker sequences and further mapping of SSR and gene markers enabled location of the QTL-containing linkage group on the short arm of chromosome 1A. Comparative mapping of the common markers for the gene for QHf.osu-1A^d in Duster and the Hf-resistance gene for QHf.osu-1A⁷⁴ in cultivar ‘2174’ showed that the two Hf resistance genes are located on the same chromosome arm 1AS, only 11.2 cM apart in genetic distance. The gene at QHf.osu-1A^d in Duster has been delimited within a 2.7 cM region.

Conclusion

Two distinct resistance genes exist on the short arm of chromosome 1A as found in the two hard red winter cultivars, 2174 and Duster. Whereas the Hf resistance gene in 2174 is likely allelic to one or more of the previously mapped resistance genes (H9, H10, H11, H16, or H17) in wheat, the gene in Duster is novel and confers a more consistent phenotype than 2174 in response to biotype GP infestation in controlled-environment assays.

Electronic supplementary material

The online version of this article (doi:10.1186/s12864-015-1297-7) contains supplementary material, which is available to authorized users.     

Identification of RAPD markers for 11 Hessian fly resistance genes in wheat

 The pyramiding of genes that confer race- or biotype-specific resistance has become increasingly attractive as a breeding strategy now that DNA-based marker-assisted selection is feasible. Our objective here was to identify DNA markers closely linked to genes in wheat (Triticum aestivum L.) that condition resistance to Hessian fly [Mayetiola destructor (Say)]. We used a set of near-isogenic wheat lines, each carrying a resistance gene at 1 of 11 loci (H3, H5, H6, H9, H10, H11, H12, H13, H14, H16 or H17) and developed by backcrossing to the Hessian fly-susceptible wheat cultivar ‘Newton’. Using genomic DNA of these 11 lines and ‘Newton’, we have identified 18 randomly amplified polymorphic DNA (RAPD) markers linked to the 11 resistance genes. Seven of these markers were identified by denaturing gradient gel electrophoresis and the others by agarose gel electrophoresis. We confirmed linkage to the Hessian fly resistance loci by cosegregation analysis in F₂ populations of 50–120 plants for each different gene. Several of the DNA markers were used to determine the presence/absence of specific Hessian fly resistance genes in resistant wheat lines that have 1 or possibly multiple genes for resistance. The use of RAPD markers presents a valuable strategy for selection of single and combined Hessian fly resistance genes in wheat improvement. Received: 20 March 1996 / Accepted: 6 September 1996     

Development of STS markers linked to Hessian fly resistance gene H6 in wheat

Hessian fly is one of the world's most destructive insect pests of wheat Triticum aestivum L. We have used the combination of near-isogenic lines (NIL) and random amplified polymorphic DNA (RAPD) analysis to screen up to 2,000 primers to identify DNA markers that are linked to gene H6 that confers resistance to biotype B of the insect. This screen produced six primers that show polymorphic fragments associated with resistance by H6. We have screened 440 F₂ individuals from a cross of the susceptible cultivar Newton and a NIL that contains H6 to verify the linkage between these markers and the resistance gene. A high-resolution genetic map was constructed based on recombination frequency. Two of the markers were tightly linked to the gene with no recombination observed, three were within 2.0 cM, and one was 11 cM from the gene. Three of the six markers were successfully converted to sequence tagged site (STS) markers. Both RAPD and STS primers were used to screen for the presence or absence of the resistance gene in wheat varieties. The identification of markers and construction of the genetic high resolution map provide the first steps toward localization of this resistance gene.     

Hessian fly resistance gene H13 is mapped to a distal cluster of resistance genes in chromosome 6DS of wheat

H13 is inherited as a major dominant resistance gene in wheat. It was previously mapped to chromosome 6DL and expresses a high level of antibiosis against Hessian fly (Hf) [Mayetiola destructor (Say)] larvae. The objective of this study was to identify tightly linked molecular markers for marker-assisted selection in wheat breeding and as a starting point toward the map-based cloning of H13. Fifty-two chromosome 6D-specific microsatellite (simple sequence repeat) markers were tested for linkage to H13 using near-isogenic lines Molly (PI 562619) and Newton-207, and a segregating population consisting of 192 F_2:3 families derived from the cross PI 372129 (Dn4) × Molly (H13). Marker Xcfd132 co-segregated with H13, and several other markers were tightly linked to H13 in the distal region of wheat chromosome 6DS. Deletion analysis assigned H13 to a small region closely proximal to the breakpoint of del6DS-6 (FL 0.99). Further evaluation and comparison of the H13-linked markers revealed that the same chromosome region may also contain H23 in KS89WGRC03, an unnamed H gene (H_WGRC4) in KS89WGRC04, the wheat curl mite resistance gene Cmc4, and a defense response gene Ppo for polyphenol oxidase. Thus, these genes comprise a cluster of arthropod resistance genes. Marker analysis also revealed that a very small intercalary chromosomal segment carrying H13 was transferred from the H13 donor parent to the wheat line Molly.Mention of commercial or proprietary product does not constitute an endorsement by the USDA.     

Association of a DNA marker with Hessian fly resistance gene H9 in wheat

The Hessian fly [Mayetiola destructor (Say)] is a major pest of wheat (Triticum aestivum L.) and genetic resistance has been used effectively over the past 30 years to protect wheat against serious damage by the fly. To-date, 25 Hessian fly resistance genes, designated H1 to H25, have been identified in wheat. With near-isogenic wheat lines differing for the presence of an individual Hessian fly resistance gene, in conjunction with random amplified polymorphic DNA (RAPD) analysis and denaturing gradient-gel electrophoresis (DGGE), we have identified a DNA marker associated with the H9 resistance gene. The H9 gene confers resistance against biotype L of the Hessian fly, the most virulent biotype. The RAPD marker cosegregates with resistance in a segregating F₂ population, remains associated with H9 resistance in a number of different T. aestivum and T. durum L. genetic backgrounds, and is readily detected by either DGGE or DNA gel-blot hybridization.Purdue University, Agric. Exp. Stn. Journal paper No. 14440     

A molecular marker associated with the H21 Hessian fly resistance gene in wheat

Near-isogenic lines in conjunction with bulked segregant analysis were used to identify a DNA marker in wheat (Triticum aestivum L.) associated with the H21 gene conferring resistance to biotype L of Hessian fly [Mayetiola destructor (Say)] larvae. Near-isogenic lines were developed by backcross introgression BC3F3:4 (Coker 797 * 4 / Hamlet) and differed by the presence or absence of H21 (on 2RL) derived from Chaupon rye (Secale cereale L.). Bulked DNA samples were prepared from near-isogenic lines and BC3F2 population individuals segregating for reaction to Hessian fly biotype L and screened for random amplified polymorphic DNA markers using 46 10mer primers. Random-amplified polymorphic DNA markers from resistant and susceptible individuals and parental lines were scored and these data were used to identify a 3 kb DNA fragment that was related to the occurrence of H21. This fragment was amplified from DNA isolated from Hamlet, a near-isogenic line carrying 2RL, and bulked DNA from resistant BC3F2 individuals, but not from the recurrent parent Coker 797 or DNA bulks from susceptible BC3F2 plants. Analysis of 111 BC3F2 segregating individuals and BC3F2:3 segregants confirmed the co-segregation of the 3 kb DNA marker with the H21 resistance gene to Hessian fly. Use of this marker could facilitate more rapid screening of plant populations for Hessian fly resistance and monitoring the introgression of H21.     

Development of wheat near-isogenic lines for powdery mildew resistance

Using three Chinese wheat cultivars, Bainong 3217, Beijing 837 and Laizhou 953, as recurrent parents, 33 near-isogenic lines (NILs) carrying 22 powdery mildew resistance genes (Pm1c, Pm2, Pm4b, Pm12, Pm13, Pm16, Pm20, Pm21, Pm23, and 13 undocumented genes) were developed. All NILs had no significant difference to their recurrent parents in the investigated traits of agronomic importance. The results of AFLP analysis indicated Jaccards genetic similarity of the NILs with their recurrent parents varied from 0.96 to 0.98, and confirmed that the NILs had high genetic similarity with their recurrent parents. The resistance to powdery mildew was stably expressed by the relevant NILs. Eleven of the NILs were tested using molecular markers linked to the resistance genes Pm1c, Pm4b, Pm13, Pm21, PmP, PmE, PmPS5A, PmPS5B, PmY39, PmY150, and PmH, and they were all found to carry the targeted genes. The potential application of these NILs in gene discovery is discussed.     

Obviation of wheat resistance to the Hessian fly through systemic induced susceptibility

Unlike most documented plant-insect interactions, Hessian fly-resistance [Mayetiola destructor (Say)] in wheat (Triticum aestivum L.) is initiated by a gene-for-gene recognition event in which plants carrying a specific R gene recognize salivary effectors encoded by a corresponding larval avirulence gene. However, dual infestation resulting from oviposition by virulent insects from 5 d before to 3 d after oviposition by avirulent insects on the same host plant, lead to systemic induced susceptibility, obviation of resistance, and ultimately the survival of both virulent and genetically avirulent progeny to adulthood. Simultaneous oviposition allowed greater survival of avirulent progeny than ovipositions separated by larger intervals. Because of the induction of plant resistance, hatch of avirulent larvae before virulent was more detrimental to rate of development than hatch of virulent before avirulent larvae. Obviation of resistance was not localized to the leaf being attacked by the virulent larvae, but also functioned across spatial distance into younger leaves. This research suggests that virulent Hessian fly larvae directly suppress the defense response of wheat, thus providing a refuge for avirulent genotypes, preserving diversity in field populations and increasing durability of deployed resistance genes.     

Characterization of new loci for Hessian fly resistance in common wheat

The discovery of several new loci for resistance to Hessian fly was reported here. QHf.uga-6AL, the late HR61 was recognized from wheat cultivar 26R61 on the distal end of 6AL with resistance to both biotypes E and vH13. It is the first gene or QTL found on this particular chromosome. QHf.uga-3DL and QHf.uga-1AL, physically assigned to the deletion bins 3DL2-0.27–0.81 and 1AL1-0.17–0.61, respectively, were detected for resistance to biotype vH13. Both QTL should represent new loci for Hessian fly resistance and the latter was detectable only in the late seedling stage when tolerance was evident. In addition, QHf.uga-6DS-C and QHf.uga-1AS had minor effect and were identified from the susceptible parent AGS 2000 for resistance to biotype E and vH13, respectively. QHf.uga-6DS-C is different from the known gene H13 on 6DS and QHf.uga-1AS is different from H9 gene cluster on 1AS. These loci also might be new components of Hessian fly resistance, although their LOD values were not highly significant. The QTL detections were all conducted on a RIL mapping population of 26R61/AGS 2000 with good genome coverage of molecular markers. The strategy used in the current study will serve as a good starting point for the discovery and mapping of resistance genes including tolerance to the pest and the closely linked markers will certainly be useful in selecting or pyramiding of these loci in breeding programs.     

Hypersensitive response of wheat to the Hessian fly

Hessian flyMayetiola destructor (Say) larvae are able to obtain food from their host plant without inflicting mechanical damage to the plant surface, apparently by secreting substances which elicit release of nutrients from plant cells surrounding the feeding site. Cells of fully susceptible plants retain their normal appearances, while in resistant plants extensive areas of cellular collapse occur. These responses indicate that hypersensitivity is the basis of wheat's resistance to the Hessian fly. The fly's feeding mechanism more closely resembles that of a pathogen than of a phytophagous insect; correspondingly, both the genetic relationship and resistance mechanism of the host plant to the parasite are of the sorts commonly associated with bacterial and fungal pathogens.     

Genetic characterization and molecular mapping of Hessian fly resistance genes derived from Aegilops tauschii in synthetic wheat

Two synthetic hexaploid wheat lines (×Aegilotriticum spp., 2n = 6x = 42, genomes AABBDD), SW8 and SW34, developed from the crosses of the durum wheat cultivar Langdon (Triticum turgidum L. var. durum, 2n = 4x = 28, genomes AABB) with two Aegilops tauschii Cosson accessions (2n = 2x = 14, genome DD), were determined to carry Hessian fly [Mayetiola destructor (Say)] resistance genes derived from the Ae. tauschii parents. SW8 was resistant to the Hessian fly biotype Great Plains (GP) and strain vH13 (virulent to H13). SW34 was resistant to biotype GP, but susceptible to strain vH13. Allelism tests indicated that resistance genes in SW8 and SW34 may be allelic to H26 and H13 or correspond to paralogs at both loci, respectively. H26 and H13 were localized to chromosome 4D and 6D, respectively, in previous studies. Molecular mapping in the present study, however, assigned the H26 locus to chromosome 3D rather than 4D. On the other hand, mapping of the resistance gene in SW34 verified the previous assignment of the H13 locus to chromosome 6D. Linkage analysis and physical mapping positioned the H26 locus to the chromosomal deletion bin 3DL3-0.81–1.00. A linkage map for each of these two resistance genes was constructed using simple sequence repeat (SSR) and target region amplification polymorphism (TRAP) markers.     

First combined resistance to Hessian fly and Sunn pest identified in synthetic hexaploid wheat

Hessian fly, Mayetiola destructor (Say), and Sunn pest, Eurygaster integriceps (Puton), are the two most damaging insect pests of wheat in North Africa, West and Central Asia. Host plant resistance is the most environmental friendly, cost-effective and practical means of controlling insect pests. Twenty synthetic hexaploid wheat lines selected as resistant to Syrian Sunn pest in 2010 were screened for resistance to Moroccan Hessian fly biotype in 2016. The Hessian fly screening was carried out in standard greenhouse flats using a randomized complete block design with three replications, with susceptible and resistant checks in every test flat. The results showed that three synthetic hexaploid wheat lines exhibited resistance to both Moroccan Hessian fly biotype and Syrian Sunn pest. This is the first record of combined resistance to these two pests in wheat. Mapping populations using these sources of resistance are being developed using double haploid techniques for subsequent genetic characterization and identification of linked molecular markers for marker assisted selection.     

Saturation and comparative mapping of the genomic region harboring Hessian fly resistance gene H26 in wheat

Resistance gene H26, derived from Aegilops tauschii Coss., is one of the most effective R genes against the Hessian fly [Mayetiola destructor (Say)], an important pest of wheat (Triticum aestivum L.). Using a limited number of PCR-based molecular markers a previous study mapped H26 to the wheat chromosomal deletion bin 3DL3-0.81-1.00. The objectives of this study were to saturate the chromosomal region harboring H26 with newly developed PCR-based markers and to investigate the collinearity of this wheat chromosomal region with rice (Oryza sativa L.) and Brachypodium distachyon genome. A population of 96 F₂ individuals segregating at the H26 gene locus was used for saturation mapping. All wheat ESTs assigned to the deletion bin 3DL3-0.81-1.00 were used to design STS (sequence tagged site) primers. The wheat ESTs mapped near H26 were further used to BLAST rice and B. distachyon genomic sequences for comparative mapping. To date, 26 newly developed STS markers have been mapped to the chromosomal region spanning the H26 locus. Two of them were mapped 1.0 cM away from the H26 locus. Comparative analysis identified genomic regions on rice chromosome 1 and Brachypodium Super contig 13 which are collinear with the genomic region spanning the H26 locus within the distal region of 3DL. The newly developed STS markers closely linked to H26 will be useful for mapped-based cloning of H26 and marker-assisted selection of this gene in wheat breeding. The results will also enhance understanding of this chromosomal region which contains several other Hessian fly resistance genes. Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture.     

Aphid (Hemiptera: Aphididae) resistance in wheat near-isogenic lines

Plant and aphid biomass, photosynthetic pigment (chlorophylls a and b and carotenoids) concentrations, and chlorophyll a/b and chlorophyll/carotenoid ratios were quantified in aphid-infested 'Tugela' near-isogenic lines (Tugela, Tugela-Dn1, Tugela-Dn2, and Tugela-Dn5). The objectives were to quantify changes of photosynthetic pigments (chlorophylls a and b, and carotenoids) caused by aphid feeding and assess resistance of wheat isolines through aphid and plant biomass analysis. Biomass of bird cherry-oat aphid, Rhopalosiphum padi (L.) (Hemiptera: Aphididae)-infested plants was lower than Russian wheat aphid, Diuraphis noxia (Mordvilko) (Hemiptera: Aphididae),- infested plants. When infested by D. noxia, all lines showed increased biomass over time, except Tugela where biomass decreased on day 12. No difference in plant biomass was detected among R. padi-infested and uninfested wheat lines. Biomass of D. noxia from Tugela (D. noxia-susceptible) was significantly higher than from plants with Diuraphis noxia-resistant Dn genes. Diuraphis noxia biomass from Tugela-Dn1 and Dn2 lines was not different from each other, but they were lower than from Tugela-Dn5. In contrast, there was no difference in R. padi biomass among wheat lines. Concentrations of chlorophylls a and b and carotenoids were significantly lower in D. noxia-infested plants compared with R. padi-infested and uninfested plants. When infested by D. noxia, chlorophyll a and b concentrations were not different among wheat lines on day 3, but they were lower in Tugela and Tugela-Dn1 than in Tugela-Dn2 and -Dn5 plants on days 6 and 12. However, no difference was detected in chlorophyll a/b or chlorophyll/carotenoid ratio among Tugela lines. The study demonstrated that Dn genes in the Tugela isolines conferred resistance to D. noxia but not to R. padi. Tugela-Dn1 was antibiotic, Tugela-Dn2 was tolerant and antibiotic, and Tugela-Dn5 was moderately antibiotic.     

High-resolution mapping of a gene conferring strong antibiosis to brown planthopper and developing resistant near-isogenic lines in 9311 background

The brown planthopper (BPH), Nilaparvata lugens Stål, is one of the most destructive pests to the rice production in the world. Thus, there is an urgency to identify new resistant genes for breeding. AC-1613 is an indica variety that has been reported to confer broad-spectrum resistance to BPH. In the present study, we found that AC-1613 exhibited strong antibiosis towards BPH insects. The body weight was significantly decreased when the insects fed on AC-1613 plants. By using BPH weight gain as an index of phenotyping, a novel dominant locus for resistance to BPH, designed as Bph30, was identified and its near-isogenic line (NIL) in 9311 background was developed. The F₂ population derived from a cross between AC-1613 and 9311 was used for mapping the gene. Through QTL scan, we located the gene on the short arm of chromosome 4 between RM16278 and RM16425, which explained 42.7% of the phenotypic variance (PEV) of BPH resistance in the F₂ population. The gene was finally located in a region flanking by simple sequence repeat (SSR) markers SSR-28 and SSR-69 through high-resolution mapping, the distance between the two markers in Nipponbare genome is 37.5 kb. In addition, SSR markers RM16294 and RM16299 tightly linked to Bph30 were applied effectively in introgressing Bph30 into elite rice cultivars. The developed NILs showed a strong antibiosis and high resistance to BPH.     

Application of STS markers for leaf rust resistance genes in near-isogenic lines of spring wheat cv. Thatcher

Sequence tagged site (STS) markers for eight resistance genes against Puccinia recondita f. sp. tritici were used to screen a set of near-isogenic lines of wheat cv. Thatcher containing in total 40 different Lr genes and their alleles. Polymerase chain reaction (PCR) analysis was carried out by using STS, SCAR and CAPS primers specific for the leaf rust resistance genes Lr1, Lr9, Lr10, Lr19, Lr24, Lr28, Lr37 and Lr47. The STS, CAPS and SCAR markers linked to resistance genes Lr9, Lr10, Lr19, Lr24, Lr37 and Lr47 were found to be reliable in diverse genetic backgrounds. The amplification product of the Lr1 gene marker was detected in the susceptible cv. Thatcher and in all of the near-isogenic lines examined except Lr2a, Lr2b, Lr2c and Lr19. The sequence analysis of PCR products amplified in lines Lr1, Lr10, Lr28 and in cv. Thatcher indicated that the near-isogenic lines and cv. Thatcher contained in the targeted chromosome region an allele that differed from the original alleles corresponding to Lr1/6*Thatcher (TLR621) and susceptible Thatcher (TH621). The amplification product specific to the STS marker of the Lr1 gene was amplified in almost all Thatcher near-isogenic lines and in cv. Thatcher because their alleles possessed primer sequences identical to the original allele TLR621. The marker for the Lr28 resistance gene was identified in line Lr28, carrying gene Lr28, and in 21 other near-isogenic lines. The sequencing of PCR products specific to Lr28 and generated in lines Lr1, Lr10 and Lr28 indicated that the lines Lr1, Lr10 and Lr28 are heterozygous in this region.     

Genotypic interaction between resistance genes in wheat and virulence genes in the Hessian fly Mayetiola destructor (Diptera: Cecidomyiidae)

The genotypic interaction between wheat resistance genes H3, H6, H7H8, H9 and virulence genes vH3, vH6, vH7vH8, vH9 of Hessian fly, Mayetiola destructor (Say), was studied in a growth chamber. Results showed that plants homozygous and heterozygous for the H3 gene expressed a high level of resistance against homozygous avirulent and heterozygous larvae carrying the vH3 virulence allele. The H7H8 genes were highly effective in the homozygous condition, but displayed a reduced level of resistance in the heterozygous condition. The H6 and H9 genes showed different levels of resistance against the reciprocal heterozygous larvae (vH6(a)vH6(A) versus vH6(A)vH6(a) and vH9(a)vH9(A) versus vH9(A)vH9(a)). Adults reared from vH6(a)vH6(A) and vH9(a)vH9(A) larvae were all males, consistent with the vH6 and vH9 X-linkage. Plants homozygous for H3, H6, H7H8, and H9 allowed for greater larval survival of heterozygous larvae, which suggests that avirulence to these resistance genes is incompletely dominant. Greater survival of homozygous avirulent larvae on heterozygous plants (H3h3, H6h6, H7h7H8h8, H9h9) suggests incomplete dominance of these wheat genes. Survival of heterozygous along with homozygous virulent larvae would reduce selection pressure for virulence in Hessian fly populations infesting fields of resistant wheat cultivars. This would be expected to slow the increase in frequency of virulence alleles that often results from deployment of resistant cultivars.