AAC Richard Soybean

AAC Richard is a food grade soybean [Glycine max (L.) Merr] cultivar with yellow hilum, high protein concentration, and good processing quality for foreign and domestic soymilk, tofu, and miso markets. It has resistance to SCN (soybean cyst nematode) (Heterodera Glycines Ichinohe). AAC Richard was developed at the Agriculture and Agri-Food Canada (AAFC) Harrow Research and Development Centre (Harrow-RDC), Harrow, Ontario and is adapted to areas of southwest Ontario with 3100 or more crop heat units and has a relative maturity of 2.3 (MG 2.3).

Fluorescent Soybean Hairy Root Construction and Its Application in the Soybean—Nematode Interaction: An Investigation

Background: The yield of soybean is limited by the soybean cyst nematode (SCN, Heterodera glycines). Soybean transformation plays a key role in gene function research but the stable genetic transformation of soybean usually takes half a year. Methods: Here, we constructed a vector, pNI-GmUbi, in an Agrobacterium rhizogenes-mediated soybean hypocotyl transformation to induce fluorescent hairy roots (FHRs). Results: We describe the operation of FHR-SCN, a fast, efficient and visual operation pathosystem to study the gene functions in the soybean-SCN interaction. With this method, FHRs were detected after 25 days in 4 cultivars (Williams 82, Zhonghuang 13, Huipizhiheidou and Peking) and at least 66.67% of the composite plants could be used to inoculate SCNs. The demographics of the SCN could be started 12 days post-SCN inoculation. Further, GmHS1pro-1 was overexpressed in the FHRs and GmHS1pro-1 provided an additional resistance in Williams 82. In addition, we found that jasmonic acid and JA-Ile increased in the transgenic soybean, implying that the resistance was mainly caused by affecting the content of JA and JA-Ile. Conclusions: In this study, we established a pathosystem, FHR-SCN, to verify the functional genes in soybeans and the SCN interaction. We also verified that GmHS1pro-1 provides additional resistance in both FHRs and transgenic soybeans, and the resistance may be caused by an increase in JA and JA-Ile contents.

Assessing direct and residual effects of cover crops on the soybean cyst nematode, Heterodera glycines

Greenhouse experiments were conducted to determine if cover crops directly decrease population densities of the soybean cyst nematode (SCN), Heterodera glycines, and/or have residual effects on reproduction of the nematode on soybean (Glycine max). Population densities of SCN were not significantly decreased by nine cover crop plants or three cover crop mixes compared to a non-planted soil control in a repeated 60-day-long greenhouse experiment. When susceptible soybeans were grown in the soils after cover crop growth, fewer SCN females formed following three annual ryegrass (Lolium multiflorum) cultivars (Bounty, King, and RootMax), the Daikon radish (Raphanus sativus var. longipinnatus) cultivar CCS779, Kodiak mustard (Brassica juncea), and a mix containing cereal rye, crimson clover (Trifolium incarnatum), plus Daikon radish (cultivars not stated) compared to following the non-planted control. In another repeated experiment, cover crops were grown for 56 days in SCN-infested soil in the greenhouse then exposed to Iowa winter conditions for 28 days to simulate winter termination of the plants. One treatment, a cover crop mix containing Bounty annual ryegrass plus Enricher Daikon radish, had a decrease in SCN population density greater than the non-planted control at the end of the experiment. Significantly fewer SCN females formed on soybeans grown following several cover crops, including the three annual ryegrass cultivars that had the suppressive residual effects in the first experiment. In summary, there were no cover crop treatments that consistently decreased SCN population densities across experiments, and only one cover crop treatment in one experiment significantly reduced SCN population densities more than a non-planted soil control. However, there was a somewhat consistent, adverse, residual effect of cover crops on reproduction of SCN on susceptible soybeans following growth of multiple cover crops.

Soybean miR159-GmMYB33 Regulatory Network Involved in Gibberellin-Modulated Resistance to Heterodera glycines

Soybean cyst nematode (SCN, Heterodera glycines) is an obligate sedentary biotroph that poses major threats to soybean production globally. Recently, multiple miRNAome studies revealed that miRNAs participate in complicated soybean-SCN interactions by regulating their target genes. However, the functional roles of miRNA and target genes regulatory network are still poorly understood. In present study, we firstly investigated the expression patterns of miR159 and targeted GmMYB33 genes. The results showed miR159-3p downregulation during SCN infection; conversely, GmMYB33 genes upregulated. Furthermore, miR159 overexpressing and silencing soybean hairy roots exhibited strong resistance and susceptibility to H. glycines, respectively. In particular, miR159-GAMYB genes are reported to be involve in GA signaling and metabolism. Therefore, we then investigated the effects of GA application on the expression of miR159-GAMYB module and the development of H. glycines. We found that GA directly controls the miR159-GAMYB module, and exogenous GA application enhanced endogenous biologically active GA1 and GA3, the abundance of miR159, lowered the expression of GmMYB33 genes and delayed the development of H. glycines. Moreover, SCN infection also results in endogenous GA content decreased in soybean roots. In summary, the soybean miR159-GmMYB33 module was directly involved in the GA-modulated soybean resistance to H. glycines.

Response of soybean genotypes from Northeast China to Heterodera glycines races 4 and 5, and characterisation of rhg1 and Rhg4 genes for soybean resistance

Summary Soybean cyst nematode (SCN, Heterodera glycines) is a devastating plant-parasitic nematode worldwide. Two SCN races, race 4 (HG Type 1.2.3.5.6.7) and race 5 (HG Type 2.5.7), with increased virulence were previously identified in Northeast China. To obtain new resistance sources to these SCN populations, the response of 62 genotypes, including 51 local varieties and breeding lines, and 11 indicator lines for SCN race and HG Type identification, were evaluated. Four new primers in the regions of two loci of GmSHMT08 (Rhg4) and GmSNAP18 (rhg1) were designed for PCR amplification and subsequent sequencing to characterise haplotypes instead of genome resequencing. Results indicated three haplotypes among 51 local genotypes; there were 26 lines in Haplotype I carrying both the rhg1-a and Rhg4-a resistant loci as in ‘Peking’, 13 lines in Haplotype II containing only the resistant rhg1-a locus but Rhg4-b susceptible loci, and 12 lines in Haplotype III with rhg1-c and Rhg4-b susceptible loci. Interestingly, there was no ‘PI 88788’-type resistance identified in Northeast China, although it accounts for 90% of sources in the USA. Two local breeding lines in Haplotype I displayed resistance to both SCN races. The resistance lines carried higher copy number (>1) of the tandem duplication at the rhg1 locus compared with susceptible lines (⩽1). The combination of the two microsatellite markers, Sat_162 on Chr 8 and 590 on Chr 18, distinguished the three haplotypes and predicted the resistance/susceptibility for SCN race 5. The knowledge of the phenotypes and molecular characteristics of 51 local breeding lines in Northeast China will accelerate the utilisation of sources for broad-based SCN resistance and marker-assisted selection.

Screening of Early Maturing Soybean Accessions for Resistance against HG Type 2.5.7 of Soybean Cyst Nematode, Heterodera glycines

Soybean cyst nematode (SCN; Heterodera glycines) is one of the devastating soybean pests worldwide, including the United States. Resistant cultivars combined with crop rotation are the primary methods for managing this nematode. SCN is known to have genetically diverse populations and can develop new virulent forms over time due to the continuous planting of cultivars derived from same source of resistance. Thus, identifying novel SCN resistant sources is of paramount importance for soybean breeding for nematode resistance. In this study, we screened 149 early maturity soybean [Glycine max (L.)] accessions for resistance to SCN HG type 2.5.7, which is one of the prevalent virulent SCN populations in North Dakota. SCN white females were extracted from individual plants of each accession after 35 days of growth in greenhouse conditions. The females were counted to determine a female index [FI = (average number of females on a tested accession/average number of females in Barnes, a susceptible soybean check) x 100]. The resistance response of each soybean accession was categorized as resistant, moderately resistant, moderately susceptible, and susceptible. Out of the soybean 149 accessions tested, only 13 were resistant in both runs of the experiments. The majority of screened soybean accessions were susceptible or moderately susceptible to the SCN HG type 2.5.7. The resistant soybean accessions identified in this study have the potential to be used in breeding SCN-resistant cultivars after further elucidation of the resistance genes or loci.

BIOLOGICAL NEMATICIDES ASSOCIATED WITH POULTRY LITTER IN THE CONTROL OF NEMATODES IN SOYBEAN AND SECOND CORN CROP

ABSTRACT Alternative management measures have been used to reduce nematode population levels in affected areas. In this perspective, the objective was to evaluate the efficiency of nematicides based on fungus and bacterium, associated with poultry litter applied via ground in the management of Pratylenchus brachyurus and Heterodera glycines in soybean and corn crops in succession. The experiment was conducted in the 2018/19 growing season in the municipality of Ipameri– GO (-17°34’55.58”S−48°12’02.35”O) in naturally infested field, arranged in a randomized block design, with 10 treatments and 4 replicates, being: T1- Witness/ control, T2- Poultry litter, T3- P. lilacinum + T. harzianum, T4- B. subtilis + B. licheniformis, T5- P. chlamydosporia, T6- Abamectina, T7- P. lilacinum + T. harzianum + Poultry litter, T8- B. subtilis + B. licheniformis + Poultry litter, T9- P. chlamydosporia + Poultry litter, T10- Abamectina + Poultry litter. The poultry litter was added in the soil 10 days before planting. The plots were composed of 6 lines of 6 m, with a spacing of 0.5 m. In the soybean crop at 45 and 90 days after sowing (DAS), the fresh weight of the roots (MFR), nematological variables and productivity were evaluated. In corn, only nematological variables were evaluated at 60 DAS. The treatments using P. lilacinum + T. harzianum and B. subtillis + B. licheniformes increased the MFR and reduced the number of P. brachyurus and H. glycines at the root. P. chlamydosporia associated with poultry litter increased in productivity in soybean plants.

Developing a Real-time PCR Assay for Direct Identification and Quantification of Soybean Cyst Nematode, Heterodera glycines, in Soil and Its Discrimination from Sugar Beet Cyst Nematode, Heterodera schachtii

The soybean cyst nematode (SCN) Heterodera glycines continues to be a major threat to soybean production worldwide. Morphological discrimination between SCN and other nematodes of the H. schachtii sensu stricto group is not only difficult and time-consuming, but also requires high expertise in nematode taxonomy. Molecular assays were developed to differentiate SCN from sugar beet cyst nematode (SBCN) and other nematodes; and to quantify SCN directly from DNA extracts of field soils. SCN and SBCN-specific quantitative real-time PCR (qPCR) primers were designed from a nematode-secreted CLAVATA gene and used for these assays. The primers were evaluated based on specificity, efficiency, and target specificity to SCN or SBCN using DNA from 20 isolates of SCN and 32 isolates of other plant-parasitic nematodes. A standard curve relating threshold cycle and log values of nematode numbers was generated from artificially infested soils and was used to quantify SCN in naturally infested field soils. There was a high correlation between the SCN numbers estimated from naturally infested field soils by conventional methods, and the numbers quantified using the SYBR Green I-based qPCR assay. The qPCR assay is highly specific and sensitive and provides improved SCN detection sensitivity down to 1 SCN egg in 20 g of soil (10 eggs/200 g soil). This assay is useful for efficient detection and quantification of SCN directly from field soil. Species-specific conventional PCR assays were also developed each for SCN and SBCN, alongside a qPCR assay that simultaneously discriminates SCN from SBCN. These assays require no expertise in nematode taxonomy and morphology, and may serve as useful diagnostic tools in research, diagnostic labs, and extension services for SCN management. Sensitive and accurate detection and quantification of SCN are essential for recommending effective management measures against SCN. We also investigated the impact of soil texture and nematode life stage on molecular quantification of SCN.