Functional Divergence of G and Its Homologous Genes for Green Pigmentation in Soybean Seeds

The degradation of chlorophyll in mature soybean seeds is closely related to the development of their yellow color. In this study, we examined G, its homologue G-like (GL), and their mutant alleles and investigated the relationship between these genes and chlorophyll accumulation in the seed coats of mature seeds. Transient expression of G and GL proteins fused with green fluorescent protein revealed that both were localized in plastids. Overexpression of G resulted in the accumulation of chlorophyll in the seed coats and cotyledons of mature seeds, indicating that high expression levels of G result in chlorophyll accumulation that exceeds its metabolism in the seeds of yellow soybean. Analysis of near isogenic lines at the G locus demonstrated a significant difference in the chlorophyll content of the seed coats and cotyledons of mature seeds when G and mutant g alleles were expressed in the d1d2 stay-green genetic background, indicating that the G protein might repress the SGR-independent degradation of chlorophyll. We examined the distribution of mutant alleles at the G and GL loci among cultivated and wild soybean germplasm. The g allele was widely distributed in cultivated soybean germplasm, except for green seed coat soybean lines, all of which contained the G allele. The gl alleles were much fewer in number than the g alleles and were mainly distributed in the genetic resources of cultivated soybean from Japan. None of the landraces and breeding lines investigated in this study were observed to contain both the g and gl alleles. Therefore, in conclusion, the mutation of the G locus alone is essential for establishing yellow soybeans, which are major current soybean breeding lines.

Patterns of Genetic Variation in a Soybean Germplasm Collection as Characterized with Genotyping-by-Sequencing

Genomic characterization is playing an increasing role in plant germplasm conservation and utilization, as it can provide higher resolution with genome-wide SNP markers than before to identify and analyze genetic variation. A genotyping-by-sequencing technique was applied to genotype 541 soybean accessions conserved at Plant Gene Resources of Canada and 30 soybean cultivars and breeding lines developed by the Ottawa soybean breeding program of Agriculture and Agri-Food Canada. The sequencing generated an average of 952,074 raw sequence reads per sample. SNP calling identified 43,891 SNPs across 20 soybean chromosomes and 69 scaffolds with variable levels of missing values. Based on 19,898 SNPs with up to 50% missing values, three distinct genetic groups were found in the assayed samples. These groups were a mixture of the samples that originated from different countries and the samples of known maturity groups. The samples that originated from Canada were clustered into all three distinct groups, but 30 Ottawa breeding lines fell into two groups only. Based on the average pairwise dissimilarity estimates, 40 samples with the most genetic distinctness were identified from three genetic groups with diverse sample origin and known maturity. Additionally, 40 samples with the highest genetic redundancy were detected and they consisted of different sample origins and maturity groups, largely from one genetic group. Moreover, some genetically duplicated samples were identified, but the overall level of genetic duplication was relatively low in the collection. These findings are useful for soybean germplasm management and utilization.

RESISTANT LEVEL OF SOYBEAN GERMPLASM AGAINST POD SUCKING BUGS (Riptortus spp.)

<p class="abstrakinggris">Increasing productivity of soybean has often been constrained by pod sucking bugs (<em>Riptortus</em> spp.) which caused a serious damage and yield losses up to 80%. Breeding for obtaining soybean variety resistant to pod suckers needs the availability of soybean germplasm resistant to the pest. The study aimed to obtain a candidate for soybean variety resistant to Riptortus spp. through the selection of 100 accessions of soybean. The study included the preparation of test plants and test insects, pest infestations, observations, and looking for a practical screening method for pod sucking pests. The experiment used a completely randomized design for two treatments (infested and non-infested <em>Riptortus</em> spp.). Cikuray variety and PI-092734 accession were used as a control. Results showed that there was a very low correlation among variables observed. Twelve soybean accessions showed a resistance to <em>Riptortus </em>spp., i.e. C7301-113AC-POP, Lokal Madiun-3549, Lokal Klungkung, ML.2974, Singgalang, Lokal Jepara, Lokal Jatim, Lokal Trenggalek, Lokal Tulungagung, Lokal Tabanan, Lokal Blitar, and Lokal Kuningan 10. These accessions were more resistant than the popular released variety such as Wilis, Grobogan, Detam 2, and Gepak Ijo. Small seed size was not a major determinant of soybean resistance to pod suckers. The addition of observational components, i.e. probing preference and oviposition, indicated that crop damage was indirectly influenced by the high frequency of probing and oviposition, although its relation to plant tolerant mechanisms still needs further investigation. Indeterminate plant types require further validation as to whether they contribute significantly to plant resistance against pod sucking insects.</p>

Molecular assessment of pathotype diversity of Phytophthora sojae in Canada highlights declining sources of resistance in soybean

The large-scale deployment of Rps (resistance to Phytophthora sojae) genes in soybean has led to the rapid evolution of the virulence profile (pathotype) of P. sojae populations. Determining the pathotypes of P. sojae isolates is important in selecting soybean germplasm carrying the proper Rps, but this process is fastidious and requires specific expertise. In this work, we used a recently developed molecular assay to assess the pathotypes of P. sojae isolates obtained throughout the provinces of Québec, Ontario and Manitoba. In preliminary assays, the molecular tool showed equivalent prediction of the pathotypes as a phenotyping assay and proved to be much faster to apply while eliminating intermediate values. Following the analysis of nearly 300 isolates, 24 different pathotypes were detected in Québec and Ontario, compared to only eight in Manitoba, where soybean culture is more recent. Pathotype 1a, 1c, 1d was predominant in Québec, while 1a, 1b, 1c, 1d, 1k was the most common in Manitoba. Overall, the results showed that 98 and 86% of the isolates carried pathotype 1a or 1c, respectively, suggesting that Rps1a and Rps1c were no longer effective in Canada. Based on the history of soybean varieties used in surveyed fields, it was found that 84% of them contained Rps genes that were no longer resistant against the pathotypes of the isolates found in the fields. While highlighting an easier and more precise option to assess pathotypes, this study presents the first pan-Canadian survey of P. sojae and stresses the importance of carefully managing the declining sources of resistance.

Development and Application of an in Vitro Method to Evaluate Anthracnose Resistance in Soybean Germplasm

Background: Anthracnose caused by Colletotrichum truncatum is a major fungal disease of soybean, especially vegetable soybean (edamame). Studies of this disease have mainly focused on resistance evaluation, but the primary methods used—in vivo inoculation of pods or plants under greenhouse or field conditions—have limitations with respect to accuracy, stability, scale, and environmental safety. Result: In this study, we developed a method for inoculating pods in vitro that entails soaking in a mycelial suspension. We optimized the crucial components, including the mycelial suspension concentration (40 to 60 mg mL-1), maturity of sampled pods (15 days after flowering), and post-inoculation incubation period (5 days). Application of the mycelial suspension by spraying rather than soaking improved the efficiency of inoculation and made large-scale evaluation possible. Using this method, we evaluated 589 soybean germplasm resources [275 cultivars (C), 233 landraces (L), and 81 wild-type accessions (W)]. This screening resulted 37 highly resistant (25 C, 11 L, and 1 W), 148 resistant (100 C, 43 L, and 5 W), 210 moderately susceptible (116 C, 77 L, and 17 W), 91 susceptible (23 C, 52 L, and 16 W), and 103 highly susceptible (11 C, 50 L, and 42 W) materials. Conclusions: The resistance levels of soybean germplasm resources were effectively distinguished by the method we developed. We thus identified many resistant cultivars but only a few resistant landraces and wild-type germplasm resources. Our results will greatly aid future research on soybean anthracnose resistance, including gene discovery, elucidation of molecular mechanisms, and the breeding of resistant cultivars.