INDIANA

PURDUE UNIVERSITY

Departments of Agronomy, Entomology, and Botany and Plant Pathology, and the USDA-ARS, Purdue University, West Lafayette, IN 47907, USA.

J. Anderson (USDA-ARS), W. Berzonsky, I. Dweikat, D. McFatridge, H. Ohm, F. Patterson, and H. Sharma (Department of Agronomy); G. Buechley, S. Goodwin (USDA-ARS), D. Huber, K. Perry, and G. Shaner (Department of Botany and Plant Pathology); R. Ratcliffe, C. Williams, S. Cambron, F. Maas, M. Jones, C. Collier, and C. Liang (USDA-ARS, Crop Production and Pest Control Research Unit (Department of Entomology)).

Production.

Farmers in Indiana seeded 850,000 acres (344,000 ha) of wheat in the fall of 1995 and harvested 720,000 acres (291,000 ha) the following spring. Average yield was 38 bu/acre (2,553 kg/ha) for a total production of 27.36 million bushels (745 million kg). The harvested area was 9 % greater than in 1995, but total production was only 69 % and yield per acre was only 37 % that of 1995. The state average yield of 38 bu/acre was the lowest since 1976. Private brands comprised 56.4 % of the acreage, compared to 64.5 % of the acreage in 1995. Among publically developed cultivars, Clark was most popular, occupying 15.9 % of the state wheat area (Data source: Indiana Agricultural Statistics Service, Purdue University).

The 1996 Indiana wheat crop was beset by several calamities. The winter was stressful, and damage to the crop was extensive. Some fields that would normally have been abandoned and planted to a spring crop were retained, because wet spring weather interfered with planting of corn and soybeans. Rainfall was frequent during the spring, which favored Stagonospora and Septoria leaf blotches and S. nodorum glume blotch. Rains continued during the flowering period throughout most of the state. Consequently, scab was widespread and severe. Stem rust was moderately severe, but developed late and was not too damaging.

Drs. Peggy Sellers and Karen Rane of the Plant and Pest Diagnostic Clinic conducted a survey for Karnal bunt by examining grain samples from virtually every county elevator in Indiana handling wheat. Results were negative. Once the Karnal bunt assays were performed, we examined these samples for scab. For the nine crop reporting districts, the average percentages of visibly scabby kernels were as follows:

Northwest 16.8 % West central 13.0 % Southwest 5.6 %

North central 25.5 % Central 8.7 % South central 6.9 %

Northeast 13.0 % East central 13.6 % Southwest 11.4 %

An informal survey of mycotoxin in wheat samples by Dr. Charles Woloshuk revealed levels of DON from 0 to 50 ppm. The FDA discourages use of wheat grain with greater than 2 ppm DON.

Natural epidemics of leaf and glume blotch and of scab were uniform in the breeding nurseries and cultivar trials. We were able to obtain excellent notes on reactions of lines to these diseases. Scab reaction was less influenced by flowering date than has been the case in previous years, so notes from this year are probably a good indication of true resistance. Incidence (percent of heads showing any symptom of scab) and severity (percent of spikelets scabbed on infected heads) were correlated (R = 0.64). However, some lines had a low incidence and high severity; whereas others had high incidence and low severity. Cultivars that show these different reactions possibly have different genes that affect scab development. We found some range in resistance to leaf blotch among cultivars available for production in Indiana, but it is not large. A few cultivars retain a green flag leaf longer than others, which should contribute to better yield and test weight.

Research on plant pathogens.

Partial resistance to leaf rust: Studies of a population derived from the cross of slow-rusting wheat CI 13227 with susceptible wheat Suwon 92 indicate that four genes with unequal effect control the partial resistance of CI 13227. A gene at one of the loci exerts a major effect on latent period. In breeding for slow-rusting resistance, slow-rusting plants may be selected from bulk populations that were carried for several generations.

We also evaluated the F7 progeny of the cross between CI 13227 and Clark. Although Clark appears to be susceptible, it displays a mixture of susceptible-type pustules and very small, resistant flecks. CI 13227 has susceptible-type pustules, but they are smaller and take longer to develop than on a fully susceptible cultivar, which accounts for its partial resistance. Many progeny of `Clark x CI 13227' cross had a greater and different kind of resistance than seen in either parent, a necrotic-fleck reaction. This resistance is evidently the result of interaction of genes from both parents, and if combined into an adapted cultivar, could give durable protection against leaf rust.

Naturally occurring strains of P. recondita were evaluated for variation in latent period on susceptible and partially resistant wheat cultivars. `Isolate x cultivar' interactions were significant. All strains had the same latent period on a susceptible cultivar, but differed in their adaptation to partially resistant cultivars. No strain completely overcame the partial resistance of any cultivar. (Shaner, Buechley, and Valezquez)

Scab: Three Midwestern and three Chinese isolates of F. graminearum were compared for virulence to wheat cultivars with different degrees of resistance. The Chinese isolates were slightly more virulent, but resistance was not cultivar-specific. Four wheat cultivars from China were consistently resistant to spread of infection within the spike, regardless of the isolate. A severe natural epidemic of scab revealed that many lines from the scab resistance breeding program had milder symptoms in the field and produced better quality grain than susceptible checks.

We have found that the cultivar Chokwang, which we originally studied as a source of partial resistance to leaf rust, has some resistance to scab. Chokwang is not as resistant as Ning 7840 or Sumai 3, but it is more resistant than any of our SRWW cultivars. The cultivar may have genes for resistance different from those in the Chinese sources, which might augment the Ning 7840 resistance.

We compared different methods of inoculation with the scab fungus on several cultivars with different degrees of resistance. Covering each inoculated head with a plastic bag resulted in greater severity than covering the head with a glassine bag. Inoculation by placing into a central floret a piece of cotton that had been dipped in a spore suspension, was as effective as injecting a spore suspension with a syringe. Injection by syringe was quicker. Regardless of inoculation method, there were significant differences among cultivars in susceptibility. Patterson was most susceptible, followed by Clark and Frontana. Sumai 3 and Ning 7840 were the most resistant. If post-inoculation high humidity was provided by only a glassine bag, there was almost no scab development on these two cultivars; if it was provided by a plastic bag (higher humidity), there was some spread of scab in the head. Soft red wheat cultivar Freedom had significantly less scab than Clark and Patterson, but more than Sumai 3 and Ning 7840.

Scab will probably need to be brought down to manageable levels through a combination of resistant cultivars and cultural practices that reduce the amount of inoculum. As part of a multistate study, we monitored the presence of spores of G. zeae on wheat heads from head emergence through flowering. The wheat heads assayed for spores were within about 60 feet of a field with abundant corn residue from 1995 on the soil surface. Spores could be detected on heads throughout this period, and subsequent scab development was severe. We do not know how far spores from corn residue can be carried by wind, and we plan to investigate this in the spring of 1997. (Bai, Shaner, and Buechley)

Foliar fungicides: Averaged over several years and locations, Tilt fungicide applied at flag leaf emergence has protected the flag leaf of wheat and increased yield an average of 5 bu/acre (336 kg/ha) and test weight an average of 1 lb/bu (13 kg/m3). A combination of Bayleton and Mancozeb applied at head emergence gave similar results. On Patterson wheat in 1996 when diseases were severe, Tilt provided more striking protection and increased yield from 67 bu/acre (4,502 kg/ha) in untreated plots to 78 bu/acre (5,241 kg/ha). (Buechley and Shaner)

Hessian fly research.

Insect surveys. Surveys were conducted to monitor Hessian fly infestations in commercial wheat fields and Purdue University or USDA wheat trials located throughout Indiana in the autumn of 1996. The incidence of Hessian fly was low in all areas where wheat was planted after the fly-free date. However, fly infestation was high in an insecticide trial established in Posey County on 25 September, 1996, approximately 2 weeks before the fly-free date for southwestern Indiana. There was an average of 37 % infested plants, and an infestation rate of two flaxseed per plant in untreated Clark control plots in this study. Adjacent wheat fields seeded approximately 2 weeks later (at or near the fly-free date) had very low incidence of Hessian fly infestation and injury. Information obtained from this trial demonstrated the susceptibility of wheat cultivars with the currently deployed Hessian fly-resistance genes to fly populations in Indiana (biotype L), and the value of delayed fall seeding as a cultural control method for the Hessian fly. (Cambron and Ratcliffe)

Biotype composition of Hessian fly populations from the mid-south and southeastern U.S. and effectiveness of resistance genes H9 to H19 to Arkansas populations. Fourteen Hessian fly populations from Arkansas, Georgia, Illinois, North Carolina, and Virginia were evaluated for biotype composition, and two Arkansas populations were tested for response to Hessian fly-resistance genes H9 to H19 in Purdue wheat lines. Hessian fly populations from Arkansas, Illinois, North Carolina, and Virginia were predominantly biotype L and virulent to all resistance genes currently deployed in soft winter wheat cultivars. Biotypes G, J, L, M, and O were identified from Hessian fly populations collected from west central Georgia, although only biotypes M and O were identified from populations sampled from southwestern Georgia. The presence of biotypes J and L in Hessian fly populations from central Georgia will reduce the effectiveness of wheat cultivars carrying the resistance gene combination H7H8 in this area. Wheat lines with single resistance gene pairs H9, H10, H13, H14, and H16 to H19 were resistant to both Arkansas Hessian fly populations, although lines with single resistance genes H11, H12, or H15 were susceptible to one or both populations. (Ratcliffe and Cambron, in cooperation with Ohm and Patterson)

Analysis of five unnamed Hessian fly-resistance genes in wheat. Triticum aestivum and T. turgidum mapping populations were constructed to localize five unnamed loci for Hessian fly resistance. Each parental line was shown to contain only a single locus for Hessian fly resistance. The five genes were rated for relative strength of resistance in heterozygotes. (Williams, Collier, Liang, and Jones, in cooperation with Ohm, Maas, and Patterson)

Gene H9. With the screening of 1,500 10-mer primers, we have identified six markers in association with gene H6 and four with gene H9. Linkage analyses were made using F2 populations of 400 progeny for H6, 1,000 progeny for H9 and 248 progeny segregating for both H6 and H9 in an effort to integrate all of the markers within the region (2 m.u.) that encompass the H6 -H9 genes. The majority of the markers (7 of the 10) were clustered within 3 cM of the two genes. Sequence analysis of these markers reveals a high degree of homology (> 90 %) between one of the markers for gene H9 and the rDNA that is located near the telomere on the long arm of chromosome 5A, suggesting that H9 is located on the long arm. (Dweikat and Ohm)

Barley yellow dwarf virus resistance research.

Strain survey. BYDV was prevalent on wheat in Tippecanoe County, Indiana, during the 1995-96 season. In plots of the cultivar Patterson, structured unbiased sampling and ELISA testing of 42 plants revealed 16 (38 %) to be infected, although symptoms were not evident. Fourteen of these plants were infected with the PAV serotype. The remaining two infected plants harbored the RPV serotype. No mixed infections were observed. The MAV, SGV, and RMV serotypes were not observed. Samplings of symptomatic wheat (n = 19) revealed 15 plants infected with either the PAV or RPV serotype; the remaining four plants were infected with both serotypes. These results contrast with observations in the 1994-95 season when both the RMV serotype and mixed infections with multiple BYDV serotypes were common. Although most of the wheat was asymptomatic, older cultivars of oats planted in adjacent fields were significantly damaged by high BYDV infection. (Perry)

BYDV-resistant germplasm. The germplasm line P29 was released. Germplasm P29 is an alien substitution line from Th. intermedium and is resistant to BYDV. The Thinopyrum chromosome that imparts BYDV resistance substitutes for chromosome 7D of wheat. The presence of the Thinopyrum chromosome was confirmed by FISH , using Th. intermedium genomic DNA as a probe. The chromosome composition of the potential translocations that were developed from Th. intermedium substitution, and addition lines studied in M4 generation, was found to be stable (2n = 42). M5 progeny testing for BYDV resistance by ELISA showed the expected results and established homozygosity for BYDV resistance. The translocations are being characterized for BYDV resistance and amount of wheatgrass DNA (J. Anderson's lab). This source of resistance is being incorporated into the breeding program. A BC1F2 population was developed from germplasm line P29 from crosses with ph mutant to induce recombinations between wheatgrass and wheat chromosomes to transfer the BYDV resistance. (Sharma and Ohm)

Molecular analysis of wheatgrass derived BYDV resistance. We have determined that an introgressed wheat line (P29), where a group 7, Th. intermedium chromosome was substituted for 7D, is moderately resistant to subgroup I BYDV strains (PAV) and completely resistant to subgroup II BYDV strains (RPV and RMV). To localize this BYDV resistance, a detailed molecular genetic analysis was made on the alien group 7 Th. intermedium chromosome contained in line P29 to determine its structural organization. Triticeae group 7 RFLP markers and rye-specific repetitive sequences used in the analysis showed that the alien chromosome in the P29 substitution line has distinguishing features. The 350-480 bp rye telomeric sequence family was present on the long arm as determined by Southern and fluorescence in situ hybridizations. However, further analysis using rye dispersed repetitive sequences indicated that this alien chromosome does not contain introgressed segments from the rye genome. The alien chromosome is homoeologous to wheat 7A and 7D as determined by RFLP analysis. The apparent colinearity with wheat group 7 chromosomes and rice chromosome 6 has allowed us to use these RFLP markers and in situ hybridization to identify translocation lines developed from g-irradiated alien substitution lines and addition lines. From this analysis, we have identified resistant translocation lines containing less than a full long arm translocation, which are being incorporated into the breeding program. The BYDV resistance locus also has been localized to the distal portion of the long arm of the group 7 Th. intermedium chromosome. (Bucholtz, Crasta, Greene, Francki, Zhu, Anderson)

Anther culture. Anthers from `wheat x wheatgrass' hybrids and backcrosses were cultured to investigate the possibility of isolating alien addition lines. Callus induction in F1 hybrids was only 0-2 % with some improvement in backcrosses. Limited data show that the chromosome numbers in the pollen-derived plants/organs of wide hybrids can be reduced to produce alien addition lines. In an effort to improve the efficiency of anther culture, co-culture of intact anthers with ovules on solid media was tried in wheat and backcrosses of the wide hybrids. Wheat genotypes differed significantly for callus induction and plantlet regeneration. Co-culture did not improve anther development in wheat, but had some beneficial effect on the ability to culture anthers of wide hybrids. (Sharma)

Septoria pathogens. We have been investigating the population biology of the Septoria pathogens to determine whether there is gene flow among populations of the pathogens infecting the major market classes of wheat in the central United States. Isolates of Mycosphaerella graminicola (S. tritici blotch), Phaeosphaeria nodorum (S. nodorum), and of the barley pathogen S. passerinii were collected in the midwestern and north central states. Twenty-nine isozyme systems were screened for polymorphisms, and eight useful systems were identified, although the level of polymorphism was low. There was very little genetic differentiation between populations of M. graminicola and P. nodorum in Indiana/Ohio and Minnesota/North Dakota, indicating either a high level of gene flow or strong selection for the isozymes identified. All three species (M. graminicola, P. nodorum, and S. passerinii) were easily distinguished by isozyme analysis alone. Analyses with RAPD primers revealed much more polymorphism. Some RAPD primers appeared to identify multiallelic loci. Bands from these loci are being cloned and sequenced for developing markers for characterizing genetic diversity in populations of these pathogens. (Goodwin)

Glume blotch. DNA from two uniformly resistant and two uniformly susceptible F7 families from a recombinant inbred population (Purdue 831800 x Purdue 871327) has been bulked and is being screened by amplifying products from RAPDs for molecular markers associated with glume blotch resistance. Bulked DNA is being screened. To date, approximately 200 primers have been screened. Putative markers will be screened again and retested for cosegregation. We are in the process of producing recombinant inbred populations for three other sources of resistance to glume blotch. F2 populations were produced by intercrossing highly susceptible and highly resistant plants originating from the same advanced Purdue lines in a field nursery evaluated in 1995. These populations will be grown and evaluated in 1997 field nurseries. (Berzonsky and Ohm)

Heterozygous translocations. Frondoso, a parent of Atlas 66 wheat, and other wheat lines originating from Brazil were examined for the presence of quadrivalents at meiosis. Six of nine lines originating from Frondoso had quadrivalents, which are indicative of a heterozygous reciprocal chromosome translocation. Frondoso, Fronteira, Atlas 50, Wakeland, and Coastal were crossed with Chinese Spring double-ditelosomic 2A and 2D lines. Meiotic analyses of the hybrids indicate they likely carry a 2A/2D translocation. Because Frondoso is a parent of Atlas 66, it is the probable source of the Atlas 66 translocation, and the detection of a 2A/2D translocation in Fronteira traces it back to an earlier Brazilian origin. Aneuploid progeny can be produced from non-cooriented segregations from these quadrivalent-forming cultivars. Genetic studies and breeding may be impacted when these cultivars are used. (Berzonsky)

Rye translocations. Four SRWW cultivars (GR 876, Freedom, Pioneer7 cultivar 2510, and Grant) having the T1RS-1BL translocation were probed for the loss of rye chromatin using rye-specific telomeric, dispersed, and centromeric sequences. In situ hybridizations and Southern hybridizations of the probes to each cultivar indicate that each maintains a complete 1RS chromosome arm despite arising from three separate breeding programs. Therefore, SRWW breeders will likely be required to deliberately manipulate the loss of 1RS chromatin to eliminate undesirable quality loci but still maintain higher yield and pest resistance loci. (Berzonsky, Francki, Anderson, and Ohm)

Personnel.

Dr. Ouafae Benlhabib from Institute of Agronomy and Veterinary, Morocco, and a collaborator on a PSTC/AID project spent 1 month at Purdue University working with H. Sharma. Dr. X. Hu joined S. Goodwin's research group in a postdoctoral position and is focusing on identifying molecular markers linked to Septoria resistance genes. Maria Manetti from Argentina began studies toward the M.S. degree with H. Ohm, developing DNA markers associated with scab resistance genes. Brad Nelson began studies toward the Ph.D. degree with H. Ohm, developing DNA markers associated with plant maturity and plant height.

Publications.

Anderson JM, Crasta O, Francki M, Bucholtz D, Sharma H, and Ohm HW. 1997. Molecular and cytogenetic analysis of barley yellow dwarf virus resistant translocation lines containing Thinopyrum intermedium chromosomal segments. Proc Plant and Animal Genome V, San Diego, CA., Jan. 12-17. p. 90.

Bai G and Shaner G. 1996. Variation in Fusarium graminearum and cultivar resistance to wheat scab. Plant Dis 80:975-979.

Benlhabib O, Hmoud R, and Sharma H. 1996. Analyse cytogenetique et androgenetique de descendants intergeneriques entre Triticum aestivum et Agropyron species. Actes IAV, Maroc 16:27-32.

Berzonsky WA. 1996. Brazilian origin and inheritance of a heterozygous reciprocal chromosome translocation in wheat (Triticum aestivum L.). Cytologia 61:253-258.

Berzonsky WA, Francki MG, Anderson JM, and Ohm HW. 1996. A cytological and molecular comparison of soft red winter wheat varieties with 1RS.1BL. Agron Abstr 88:73.

Berzonsky WA and Ohm HW. 1997. Breeding cereal small grains for value-added uses. ASA monograph chapter. In press.

Buechley G and Shaner G. 1996. Control of bunt and loose smut of wheat with fungicidal seed treatments, 1995. Fungicide and Nematicide Tests 51:307.

Cambron SE, Ohm HW, Ratcliffe RH, and Patterson FL. 1996. A second gene for resistance to Hessian fly in Iumillo Wheat. Crop Sci 36:1099-1101.

Cebert E, Ohm H, Patterson F, Ratcliffe R, and Cambron S. 1996. Genetic analysis of Hessian fly resistance in durum wheat. Agron Abstr 88:88.

Crasta O, Francki M, Greene A, Bucholtz D, Sharma H, Ohm H, and Anderson J. 1996. Molecular characterization of Thinopyrum chromatin in wheat toward targeted mapping of BYDV resistance. 2nd Inter Crop Sci Cong, New Delhi, 17-24 November, 1996.

Desjardins AE, Proctor RH, Bai G, McCormick SP, Shaner G, Buechley G, and Hohn T. 1996. Reduced virulence of trichothecene-nonproducing mutants of Gibberella zeae in wheat field tests. Mol Plant-Microbe Inter 9:775-781.

Drake DR and Ohm HW. 1996. Inheritance of head scab resistance in wheat. Agron Abstr.

Dweikat I, Ohm H, Patterson F, and Cambron S. 1997. Identification of RAPD markers for 11 Hessian fly resistance genes in wheat. Theor Appl Genet (in press).

Dweikat I, Zhang W, Ohm H, Cambron S, and Ratcliffe R. 1996. Construction of a high resolution map for Hessian fly resistance genes H6 and H9 in wheat. Proc Plant Genome IV, 15-19 January, San Diego, CA. (Abstract).

Gyulai G, Dweikat I, Janovsky J, Ohm H, Kiss E, Sharma H, and Heszky L. 1996. Analysis of Agropyron, Bromus and Agropyron x Bromus by ISSR-PCR. EUCARPIA Cong. Poland, 6-11 October, 1996.

Hodges TK, Aldemita R, Kononowicz-Hodges H, Macdonald B and Anderson JM. 1996. Agrobacterium-mediated transformation of japonica and indica rice varieties. Workshop on In Vitro Manipulation of Wheat and Small Grains, World Cong In Vitro Biol, San Francisco, 22-27 June, 1996.

Hu XY, Bostwick D, Sharma H, Ohm H, and Shaner G. 1996. Chromosome and chromosomal arm locations of genes for resistance to Septoria glume blotch in wheat cultivar Cotipora. Euphytica 91:251-257.

Hu XY, Ohm HW, and Dweikat I. 1997. Identification of RAPD markers linked to PM1 for resistance to powdery mildew in wheat. Theor Appl Genet (in press).

Lehman JS and Shaner G. 1996. Genetic variation in latent period among isolates of Puccinia recondita f. sp. tritici on partially resistant wheat cultivars. Phytopathology 86:633-641.

Ohm HW and Berzonsky WA. 1996. Screening a recombinant inbred population for resistance to glume blotch. Agron Abstr 88:73.

Ohm HW, Ratcliffe RH, Patterson FL, and Cambron SE. 1997. Resistance to Hessian fly conditioned by genes H19 and proposed gene H27 of durum wheat line PI422297. Crop Sci 37:113-115.

Patterson FL, Ohm HW, Johnson JW, and Wickersham DS. 1996. Registration of five wheat pollen fertility restorer germplasm lines: PR143, PR189, PR267, PR270, and PR302. Crop Sci 36:1424.

Ratcliffe RH, Ohm HW, Patterson FL, Cambron SE, and Safranski GG. 1996. Response of resistance genes H9-H19 in wheat to Hessian fly (Diptera: Cecidomyiidae) laboratory biotypes and field populations from the eastern United States. J Econ Entomol 89:1309-1317.

Ratcliffe RH and Hatchett JH. 1997. Biology and genetics of the Hessian fly and resistance in wheat. In: New Developments in Entomology (Bondari K ed). Research Signpost, Scientific Information Guild, Trivandrum, India. In press.

Ratcliffe RH, Ohm HW, Patterson FL, and Cambron SE. 1997. Biotype composition of Hessian fly (Diptera: Cecidomiidae) populations from Arkansas, Georgia, Illinois, North Carolina and Virginia. J Entomol Sci 32:in press.

Shaner G and Buechley G. 1996. Effect of foliar fungicides on control of wheat diseases, 1995. Fungicide and Nematicide Tests 51:231.

Sharma H. 1996. Maintenance of haploid genome of Agropyron junceum in wheat. Wheat Info Serv 83:21-24.

Sharma H, Ohm H, and Perry K. 1997. Registration of wheat germplasm line P29 with BYDV resistance. Crop Sci 37:In press.

Sharma H, Jekkel Z, Benlhabib O, and Ohm H. 1996. Effect of concomitant ovule culture on anther culturability in wheat and wheatgrasses. World Cong In Vitro Biol, San Francisco, 22-27 June, 1996. p. 84A.

Sharma H, Benlhabib O, Jekkel Z, and Ohm H. 1996. Anther culture and concomitant ovule culture of wheat x wheatgrass hybrids and wheat. Agron Abstr 88:160.

Yang X, Ohm H, and Dweikat I. 1996. Identification of RAPD markers linked to a gene for resistance to Septoria tritici in wheat. Agron Abstr 88:169.

Zhang G and Goodwin SB. 1996. Enzyme polymorphism among isolates of Mycosphaerella graminicola (anamorph Septoria tritici) and Septoria passerinii. Phytopathology 86:S90.

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