Senyu Chen

Soybean Cyst Nematode Population Development and Associated Soybean Yields of Resistant and Susceptible Cultivars in Minnesota

The soybean cyst nematode (SCN), Heterodera glycines, is a major soybean yield-limiting factor, and the use of resistant cultivars is one of the most effective means to manage the nematode. During the past decade, a number of resistant cultivars in maturity groups I and II have been developed and made available to growers. A total of 47 resistant cultivars and nine susceptible cultivars were evaluated at 15 SCN-infested field sites and two noninfested sites during 1996 to 1998 in Minnesota. As expected, more nematodes developed on susceptible cultivars than on resistant cultivars. Egg density on susceptible cultivars increased by 1.9- to 10.6-fold during the growing season at 12 sites and did not change at the other three sites. Average egg density decreased over time for resistant cultivars at all sites, except where the initial egg density was low (≤455 eggs per 100 cm3 soil). Nematode reproduction factors (Rf = egg density at harvest/egg density at planting) for individual resistant and susceptible cultivars were highly consistent across the eight sites where initial SCN density was more than 1,000 eggs per 100 cm3 soil. Resistance, however, varied among the cultivars, with the average Rf of individual resistant cultivars across the sites ranging from 0.3 to 1.7. Resistant cultivars produced an average yield of 3,082 kg/ha compared with 2,497 kg/ha by susceptible cultivars at eight of 10 sites where egg density at planting was greater than 700 eggs per 100 cm3 soil. In contrast, no difference in yield was observed between resistant and susceptible cultivars at sites where egg density at planting was lower than 500 eggs per 100 cm3 soil. Yield differences between resistant and susceptible cultivars increased with increasing initial SCN egg density. In six fields infested with initial densities of more than 5,000 eggs per 100 cm3 soil, resistant cultivars produced 28.4% (676 kg/ha) more yield on average than susceptible cultivars. Soybean yield increased when cultivars with increasing resistance to the SCN (lower Rf or females formed on roots) were grown in fields infested with SCN. Average relative yield (yield of a cultivar/average yield of all resistant cultivars at a site) of individual resistant cultivars across all SCN-infested sites ranged from 0.76 to 1.10. Yield consistency of soybean cultivars was low among the different sites, indicating that many other factors affected yield. Our results suggest growing resistant cultivars is an effective method to manage SCN in Minnesota while minimizing yield loss due to SCN.

Screening Isolates of Hirsutella Species for Biocontrol of Heterodera glycines

The fungi, Hirsutella rhossiliensis and Hirsutella minnesotensis, are two endoparasites of second-stage juveniles (J2) of the soybean cyst nematode (SCN), Heterodera glycines. The objective of this study was to screen for effective fungal isolates to control the nematode in laboratory and greenhouse assays. A total of 93 isolates of H. rhossiliensis and 25 isolates of H. minnesotensis were evaluated for parasitism of SCN J2 on cornmeal agar. Percentage of SCN J2 parasitized by the fungi varied among the fungal isolates. Most H. rhossiliensis isolates parasitized a high percentage of J2. The isolates of H. rhossiliensis obtained from bacteria-feeding nematodes, however, generally did not parasitize J2 on agar. H. minnesotensis parasitized fewer J2 on agar than did H. rhossiliensis . Forty isolates of H. rhossiliensis and four isolates of H. minnesotensis that parasitized a high percentage of J2 on agar were evaluated for their biocontrol potential in soil treated with microwave heating. Most isolates selected from the agar assay also parasitized a high percentage of J2 in the soil but there was variation among isolates. Pathogenicity of 14 isolates of H. rhossiliensis and four isolates of H. minnesotensis to the SCN was also investigated in the greenhouse using untreated field soil. Most isolates of H. rhossiliensis reduced SCN population density and increased plant growth when compared with 1% corn-grits control (culture media). One isolate (OWVT-1) of H. rhossiliensis reduced the SCN egg density by 95% and J2 density by 98% when compared with the control. Isolates of H. minnesotensis, however, neither reduced SCN density nor increased plant growth in the greenhouse.

Hirsutella minnesotensis sp. nov., a new pathogen of the soybean cyst nematode'

Hirsutella minnesotensis, a new fungal spe- cies, was isolated and described from second-stage ju- veniles of the soybean cyst nematode, Heterodera gly- cines. The fungus can be distinguished from other species of Hirsutella by the strongly swollen base of conidiogenous cells and verrucose, globose conidia. The fungus is similar to Hirsutella thompsonii in col- ony morphology and conidium characteristics, but differs from Hi. thompsonii in that the new species has a strongly swollen base, a conspicuous swelling near the apex of conidiogenous cells, and larger co- nidia. The fungus is a pathogen of He. glycines and may have potential as a biological control agent.

Potential of Association Mapping and Genomic Selection to Explore PI 88788 Derived Soybean Cyst Nematode Resistance

The potential of association mapping (AM) and genomic selection (GS) has not yet been explored for investigating resistance to soybean cyst nematode (SCN), the most destructive pest affecting soybean. We genotyped 282 representative accessions from the University of Minnesota soybean breeding program using a genome‐wide panel of 1536 single nucleotide polymorphism (SNP) markers and evaluated plant responses to SCN HG type 0. After adjusting for population structure, AM detected significant signals at two loci corresponding to rhg1 and FGAM1 plus a third locus located at the opposite end of chromosome 18. Our analysis also identified a discontinuous long‐range haplotype of over 600 kb around rhg1 locus associated with resistance to SCN HG type 0. The same phenotypic and genotypic datasets were then used to access GS accuracy for prediction of SCN resistance in the presence of major genes through a sixfold cross‐validation study. Genomic selection using the full marker set produced average prediction accuracy ranging from 0.59 to 0.67 for SCN resistance, significantly more accurate than marker‐assisted selection (MAS) strategies using two rhg1‐associated DNA makers. Reducing the number of markers to 288 SNPs in the GS training population had little effect on genomic prediction accuracy. This study demonstrates that AM can be an effective genomic tool for identifying genes of interest in diverse germplasm. The results also indicate that improved MAS and GS can enhance breeding efficiency for SCN resistance in existing soybean improvement programs.

Nutritional Requirements of the Nematophagous Fungus Hirsutella rhossiliensis

Six natural media were examined for growth and sporulation of six isolates of the nematophagous fungus Hirsutella rhossiliensis , using solid and/or liquid culture. Twenty carbohydrates, 19 nitrogen (N) compounds, and nine vitamins were also tested for their effects on growth, sporulation, and spore germination of a further three isolates (ATCC46487, OWVT-1 and JA16-1). Variations in nutritional requirements existed among the fungal isolates. In general, V-8 juice agar (VA), cornmeal agar and potato dextrose agar were good media for growth, and malt extract agar, VA and yeast dextrose agar were good for sporulation of all six isolates. Glycogen was the best and sucrose, inulin, D- ( + ) - trehalose and soluble starch were also good carbon (C) sources for growth and spore germination of the three isolates ATCC46487, OWVT-1 and JA16-1 in both liquid and solid culture. None of the isolates utilized D- ( + )xylose as a C source. L- sorbose, D- ribose, citric acid and D- fructose were poor for growth of all isolates. The best C source for sporulation was D- ( + )-trehalose for ATCC46487, D- sorbitol for OWVT-1 and D- ( + )-cellobiose for JA16-1. Casein was the best N source for growth of ATCC46487 and OWVT-1, while peptone was best for JA16-1. L- asparagine, L- proline, and peptone were also good for growth of all three isolates. L - cystine was not utilized by H. rhossiliensis and DL- methionine inhibited growth of all isolates. Spore germination of all isolates was well supported by most N compounds examined but was inhibited by L- cystine. No significant difference in sporulation of ATCC46487 was observed among the N sources. DL- threonine was the best N source for spore production by OWVT-1 and L- phenylalanine was best for JA16-1. Vitamins generally enhanced fungal growth and sporulation, with thiamine having the greatest influence. Excluding some vitamins individually from the medium containing all other test vitamins sometimes increased growth and/or sporulation of certain isolates.

Suppression of Heterodera glycines in soils from fields with long-term soybean monoculture

Abstract Experiments were conducted in a greenhouse to evaluate soils for their degree of suppressiveness against the soybean cyst nematode, Heterodera glycines. The soils were collected in 2003 and 2004 from two fields with long-term soybean monoculture and one field with corn/soybean annual rotation. The soils were autoclaved, treated with formalin, or untreated. Both formalin and autoclave treatments removed suppressive factors and increased the nematode egg population density as compared with untreated soil. The mixture of 10% untreated soil with 90% treated soil resulted in similar nematode population densities as in the untreated soils collected from the two monoculture fields, indicating the suppressive factors were biological and can be transferred. The soils from the monoculture fields appeared to be more suppressive than the soil from the corn/soybean rotation. Hirsutella rhossiliensis was observed in all three soils, and parasitised a large percentage of second-stage juveniles of H. glycines in the monoculture soils in 2004. The fungus may be a major factor for the suppression of the nematode population, but in this study other biological factors cannot be ruled out.

Nutritional requirements of Pochonia chlamydospora and ARF18, fungal parasites of nematode eggs.

Twenty carbohydrates (C), 18 nitrogen compounds (N), and 9 vitamins were examined for their effects on the growth and conidiation of the nematode-egg-parasitic fungi Arkansas Fungus 18 (ARF18, isolate 908) and Pochonia chlamydospora var. chlamydospora in solid and liquid cultures. Glycogen was the best, and inulin, D-(+)-galactose, and soluble starch were good C sources for the growth of ARF18 in both solid and liquid cultures. ARF18 could not utilize alpha-cellulose in liquid; alpha-lactose and D-mannitol in solid; and D-(+)-xylose, L-(-)-sorbose, and D-(-)-ribose in both solid and liquid cultures. Casein was the best N source for ARF18 growth in both solid and liquid cultures and L-aspartic acid, DL-glutamic acid, peptone, and L-histidine were good in solid culture. ARF18 could not utilize L-cystine and L-tyrosine in solid culture, and L-cystine, DL-methionine, peptone, L-proline, and ammonium nitrate in liquid culture. Supplement of vitamins appeared to be unnecessary for ARF18 to grow. For P. chlamydospora var. chlamydospora growth, all test C sources, except L-(-)-sorbose, alpha-cellulose, citric acid, and D-(+)-glucose, were good in both solid and liquid cultures. Most N compounds were good for P. chlamydospora var. chlamydospora growth with casein and peptone the best. Vitamins had limited effect on P. chlamydospora var. chlamydospora growth. P. chlamydospora var. chlamydospora conidiation was well supported by D-(-)-ribose, D-(-)-fructose, melibiose, and D-(+)-galactose as C sources and by L-aspartic acid, DL-glutamic acid, and L-arginine as N sources. Excluding myo-inositol from the medium containing all other test vitamins increased P. chlamydospora var. chlamydospora conidiation, while excluding pyridoxine appeared to reduce its conidiation.

Response of Heterodera glycines and Soybean Cultivar to Tillage and Row Spacing

Heterodera glycines, commonly known as the soybean cyst nematode (SCN), has become a major factor in soybean production in the Midwest United States. The influence of five tillage treatments and two treatments of row spacing on SCN population dynamics and yield of SCN-resistant and -susceptible soybean cultivars was investigated in a corn-soybean rotation system in southern Minnesota from 1993 to 1996. No effects of tillage and row spacing were observed on nematode population density. As expected, the susceptible cultivar Sturdy consistently supported higher nematode densities than did the resistant cultivar Bell in 1993 to 1995 and Freeborn in 1996. Nematode reproduction varied among years. Predicted nematode density at equilibrium was 3,800, 13,000, 12,000, and 27,000 eggs per 100 cm3 of soil in plots with the susceptible cultivar and 480, 240, 430, and 700 eggs per 100 cm3 of soil in plots with the resistant cultivars in 1993, 1994, 1995, and 1996, respectively. The effects of tillage and row spacing on soybean yield were inconsistent. The resistant cultivars yielded 653, 195, and 435 kg/ha more (P < 0.05) than the susceptible cultivar in 1994, 1995, and 1996, respectively, but no yield difference between susceptible and resistant cultivars was observed in 1993. Planting resistant cv. Bell increased the yield of the following susceptible cv. Sturdy compared with continual planting of the susceptible cultivar. A sequence with continued resistant cultivar or cultivars, however, produced a higher overall yield and lower nematode density at the end of the 4-year rotation cycle than any sequence in which the susceptible cultivar was included. Yield of resistant and susceptible cultivars was negatively related to the SCN initial population density.

Suppression of Meloidogyne hapla populations by Hirsutella minnesotensis

Abstract The effects of the endoparasitic fungus Hirsutella minnesotensis on populations of Meloidogyne hapla from Michigan (MI), Rhode Island (RI), Connecticut (CT), Lyndonville, New York (NYL), Geneva, New York (NYG), and Wisconsin (WI) were studied in the greenhouse. Twenty-day-old tomato (cv. Rutgers) seedlings were inoculated with either 0 or 600 eggs of each nematode population mixed with either 0, 0.02, or 0.1 g of fresh H. minnesotensis mycelium 0.1 L−1 of soil in pots containing 0.5 L of soil, and maintained at 25±2°C for 2 months. No effect of the fungal treatments and nematode treatments on tomato plant heights and shoot dry weights was observed. While all M. hapla populations were suppressed by H. minnesotensis, the degree to which each population was affected varied slightly. Across fungal treatments and nematode populations, the fungus reduced total number of nematodes in roots by 61–98%, with the highest for NYG and RI, intermediate for NYL and CT, and lowest for MI and WI populations. The study demonstrated that H. minnesotensis may be used as a potential suppressor of M. hapla in vegetable production systems in the Great Lakes Region.

Mycofloras in cysts, females, and eggs of the soybean cyst nematode in Minnesota

Abstract During 1996–1997, mycofloras in cysts, females, and eggs of the soybean cyst nematode (SCN), Heterodera glycines , were investigated in 45 fields from 26 counties across southern Minnesota, USA. A total of 4500 cysts, 4500 females, and 45,000 eggs were examined, and 62 fungal species were isolated and identified from the nematode. Fungi colonized 55% (29–89%) of cysts with Cylindrocarpon destructans , Fusarium solani , Pyrenochaeta terrestris , Fusarium oxysporum and Cylindrocarpon olidum the most common. The frequency of fungi encountered in cysts was positively correlated with egg density in fields. Only 3.4% of females were colonized by fungi with Oidiodendron cerealis , F. solani , F. oxysporum , C. destructans , and P. terrestris the most common. Only 1% of eggs yielded fungi with C. destructans , F. solani , P. terrestris , Exophiala pisciphila , and F. oxysporum the most common. Average fungal egg-parasitic index (EPI at a scale from 0 to 10 to measure fungal parasitism of eggs) in the fungus-colonized cysts was 2.0, and average EPI in females was 1.8. The EPI was positively correlated with percentage of cysts colonized by fungi and egg density in the fields. The Simpson’s diversity index (at a scale from 0 to 1) was low (average 0.25), indicating that fungal community in the cysts was highly diversified and no single species was dominant. Average similarity index (at a scale from 0 to 1) for mycofloras in cysts between fields in Minnesota was 0.53 (range 0.13–0.85). The study demonstrated that the mycofloras in the cysts and females from the Minnesota fields might have limited antagonistic effect on the SCN population.