Association mapping for partial resistance to Phytophthora sojae in soybean (Glycine max (L.) Merr.)

Association mapping is a powerful high-resolution mapping tool for complex traits. The objective of this study was to identify QTLs for partial resistance to Phytophthora sojae. In this study, we evaluated a total of 214 soybean accessions by the hypocotyl inoculation method, and 175 were susceptible. The 175 susceptible accessions were then evaluated for P. sojae partial resistance using slant board assays. The 175 accessions were screened with 138 SSR markers that generated 730 SSR alleles. A subset of 495 SSR loci with minor allele frequency (MAF) ≥ 0.05 was used for association mapping by the Tassel general linear model (GLM) and mixed linear model (MLM) program. This soybean population could be divided into two subpopulations and no or weak relatedness was detected between pairwise accessions. Four SSR alleles, Satt634-133, Satt634-149, Sat_222-168 and Satt301-190, associated with partial resistance to P. sojae were detected by both GLM and MLM methods. Of these identified markers, one marker, Satt301, was located in regions where P. sojae resistance QTL have been previously mapped using linkage analysis. The identified markers will help to understand the genetic basis of partial resistance, and facilitate future marker-assistant selection aimed to improve resistance to P. sojae and reduce disease-related mortality in soybean.     

Genetic mapping of a novel gene for soybean aphid resistance in soybean (Glycine max [L.] Merr.) line P203 from China

The soybean aphid (SA: Aphis glycines Matsumura) is a worldwide pest of soybean (Glycine max [L.] Merr.). The objectives of this study were to identify the type of aphid resistance and the resistance phenotype in soybean line ‘P203’, and to map the relative position of the gene involved. Compared with cultivars ‘P746’ and ‘Dongnong 47’, P203 was demonstrated to possess antixenosis resistance. P203 prevented aphids from reproducing in a choice test, but the resistance level decreased significantly in a no-choice test at 11 and 21 days after infestation. Analysis of 273 Dongnong 47/P203 F₂ plants and confirmed using 260 F_2:3 families revealed that a single dominant gene from P203 was positioned between marker loci Sat_377 and Satt409 on chromosome 8. The gene was further mapped to a 1.57 Mb interval flanked by marker loci BARCSOYSSR_08_1451 and BARCSOYSSR_08_1527. We developed five new SSR markers in the target interval and the resistance locus mapped between new markers SSR_08_75 and SSR_08_88 with the genetic distance of 1.1 and 1.0 cM corresponding to a physical distance of 192 kb on the Williams 82 8X draft genome assembly (Glyma1.01). A single serine/threonine protein kinase gene is present in this region, suggesting that the SA resistance mechanism in P203 may be different from those previously reported. Therefore, the resistance gene could very well be novel, and could be valuable in soybean aphid resistance breeding programs.     

RFLP mapping using near-isogenic lines in the soybean [Glycine max (L.) Merr.]

Summary A molecular marker analysis of a near-isogenic line (NIL), its donor parent (DP), and its recurrent parent (RP) can provide information about linkages between molecular markers and a conventional marker introgressed into the NIL. If the DP and RP possess different alleles for a given molecular marker, and if the NIL possesses the same allele as the DP, then it is reasonable to presume a linkage between that molecular marker and the introgressed marker. In this study, we examined the utility of RFLPs as molecular markers for the NIL genemapping approach. The allelic status of fifteen RFLP loci was determined in 116 soybean RP/NIL/DP line sets; 66 of the Clark RP type and 50 of the Harosoy RP type. Of the 1740 possible allelic comparisons (116 NILs x 15 RFLP loci), 1638 were tested and 462 (33.9%) of those were informative (i.e., the RP and DP had different RFLP alleles). In 15 (3.2%) of these 462 cases the NIL possessed the DP-derived RFLP allele, leading to a presumption of linkage between the RFLP locus and the introgressed conventional marker locus. Two presumptive linkages, pK-3 — and pK-472 — Lf _i, were subsequently confirmed by cosegregation linkage analysis. Although not yet confirmed, two other associations, pk-7 ab and pK-229 — y ₉ seemed to be plausible linkages, primarily because the pk-7 — ab association was detected in two independently derived NILs and both markers of the pK-229 — y ₉ association were known to be linked to Pb. The data obtained in this investigation indicated that RFLP loci were useful molecular markers for the NIL gene-mapping technique.Published as Paper no. 9101, Journal Series, Nebraska Agric. Res. Div. Project no. 12-091. Research partially funded by a grant from the Nebraska Soybean Development, Utilization, and Marketing Board     

Genetics and cytology of a new genic male-sterile soybean [Glycine max (L.) Merr.]

 Genetic and cytological studies were conducted with a new male-sterile, female-fertile soybean [Glycine max (L.) Merr.] mutant. This mutant was completely male sterile and was inherited as a single-recessive gene. No differences in female or male gamete transmission of the recessive allele were observed between reciprocal cross-pollinations in the F₁ or F₂ generations. This mutant was not allelic to any previously identified soybean genic male-sterile mutants: ms1, ms2, ms3, ms4, ms5, or ms6. No linkage was detected between sterility and flower color (W1 locus), or between sterility and pubescence color (T1 locus). Light microscopic and cytological observations of microsporogenesis in fertile and sterile anthers were conducted. The structure of microspore mother cells (MMC) in male-sterile plants was identical to the MMCs in male-fertile plants. Enzyme extraction analyses showed that there was no callase activity in male-sterile anthers, and this suggests that sterility was caused by retention of the callose walls, which normally are degraded around tetrads at the late tetrad stage. The tapetum from male-sterile anthers also showed abnormalities at the tetrad stage and later stages, which were expressed by an unusual formation of vacuoles, and by accumulation of densely staining material. At maturity, anthers from sterile plants were devoid of pollen grains. Received: 13 May 1996 / Revision accepted: 19 August 1996     

Cryopreservation of the SB-M photosynthetic soybean [Glycine max (L.) Merr.] suspension culture

Summary The photosynthetic cell suspension culture of soybean [Glycine max (L.) Merr. cv. Corsoy] (SB-M) was successfully cryopreserved in liquid nitrogen using a preculture and controlled freezing to −40° C (two-step) freezing method. The effective method included a preculture treatment with gradually increasing levels of sorbitol added to the 3% sucrose already present in the medium. The cells were then placed in a cryoprotectant solution [10% DMSO (dimethylsulfoxide) and 9.1% sorbitol, or 10% DMSO and 8% sucrose], incubated for 30 min at 0° C, cooled at a rate of 1° C/min to −40° C, held at −40° C for 1 h, and then immersed directly into liquid nitrogen. The cells were thawed at 40° C and then immediately placed in liquid culture medium. The cell viabilities immediately after thawing were 75% or higher in all cases where cell growth resumed. The original growth rate and chlorophyll level of the cells was recovered within 40 to 47 d. If the sorbitol level was not high enough or the preculture period too short, growing cultures could not be recovered. Likewise, survival was not attained with cryoprotectant mixtures consisting of 15% DMSO, 15% glycerol, and 9.1% sucrose or 15% glycerol and 8% sucrose. The successful method was reproducible, thus allowing long-term storage of this and certain other unique photosynthetic suspension cultures in liquid nitrogen.     

Cloning, mapping, and analyses of expression of the Em-like gene family in soybean [Glycine max (L). Merr.]

 The entire Em-like Group-1 late embryogenesis abundant (Lea) gene family from soybean was cloned and characterized. The five Group-1 Lea genes (Sle1-5) were divided into two classes based on sequence identity. Sle1-4 were genetically mapped to four different linkage groups. Nucleotide sequencing indicated that Sle1, Sle2, Sle3, and Sle5 encode polypeptides differing primarily by the presence of a repeated 20-amino acid motif. Sle1 and Sle5 were shown by Northern analysis to be expressed in developing embryos weeks earlier than Sle2 and Sle3. Sle4 was shown to be a pseudogene. Maximal levels of mRNA for all functional Sle genes accumulated in maturation-phase seeds, before significant desiccation had occurred, and declined rapidly upon seed imbibition. Desiccation did not induce Sle expression in seeds or vegetative tissue. Sle expression was confined to embryo tissues and Sle mRNA accumulated at similar levels in both the embryo axis and in the cotyledons. Received: 25 September 1996 / Accepted: 27 September 1996     

Factors underlying genotypic differences in the induction of photosynthesis in soybean [Glycine max (L.) Merr.]

Crop leaves are subject to continually changing light levels in the field. Photosynthetic efficiency of a crop canopy and productivity will depend significantly on how quickly a leaf can acclimate to a change. One measure of speed of response is the rate of photosynthesis increase toward its steady state on transition from low to high light. This rate was measured for seven genotypes of soybean [Glycine max (L.) Merr.]. After 10 min of illumination, cultivar ‘UA4805’ (UA) had achieved a leaf photosynthetic rate (P_n) of 23.2 μmol · m^?2 · s^?1, close to its steady‐state rate, while the slowest cultivar ‘Tachinagaha’ (Tc) had only reached 13.0 μmol · m^?2 · s^?1 and was still many minutes from obtaining steady state. This difference was further investigated by examining induction at a range of carbon dioxide concentrations. Applying a biochemical model of limitations to photosynthesis to the responses of P_n to intercellular CO₂ concentration (C_i), it was found that the speed of apparent in vivo activation of ribulose‐1:5‐bisphosphate carboxylase/oxygenase (Rubisco) was responsible for this difference. Sequence analysis of the Rubisco activase gene revealed single nucleotide polymorphisms that could relate to this difference. The results show a potential route for selection of cultivars with increased photosynthetic efficiency in fluctuating light.     

Fertile progeny of a hybridization between soybean [Glycine max (L.) Merr.] and G. tomentella Hayata

Summary A colchicine-doubled F₁ hybrid (2n=118) of a cross between PI 360841 (Glycine max) (2n=40) x PI 378708 (G. tomentella) (2n=78), propagated by shoot cuttings since January 1984, produced approximately 100 F₂ seed during October 1988. One-fourth of the F₂ plants or their F₃ progeny have been analyzed for chromosome number, pollen viability, pubescence tip morphology, seed coat color, and isoenzyme variation. Without exception, all plants evaluated possessed the chromosome number of the G. max parent (2n=40). Most F₂ plants demonstrated a high level of fertility, although 2 of 24 plants had low pollen viability and had large numbers of fleshy pods. One F₂ plant possessed sharp pubescence tip morphology, whereas all others were blunt-tipped. All evaluated F₂ and F₃ plants expressed the malate dehydrogenase and diaphorase isoenzyme patterns of the G. max parent and the endopeptidase isoenzyme pattern of the G. tomentella parent. Mobility variants were observed among progeny for the isoenzymes phosphoglucomutase, aconitase, and phosphoglucoisomerase. This study suggests that the G. Tomentella chromosome complement has been eliminated after genetic exchange and/or modification has taken place between the genomes.Journal Paper No. J-13776 of the Iowa Agriculture and Home Economics Experiment Station, Ames, IA, USA, Project 2763     

Photosynthesis and photorespiration in soybean [Glycine max (L.) Merr.] chronically exposed to elevated carbon dioxide and ozone

The effects of elevated carbon dioxide (CO₂ and ozone (O₃) onsoybean (Glycine max (L.) Merr.] photosynthesis and photorespiration-relatedparameters were determined periodically during the growing seasonby measurements of gas exchange, photorespiratory enzyme activitiesand amino acid levels. Plants were treated in open-top fieldchambers from emergence to harvest maturity with seasonal meanconcentrations of either 364 or 726 µmol mol^–1 CO₂in combination with either 19 or 73 nmol mol^–1 O₃ (12h daily averages). On average at growth CO₂ concentrations,net photosynthesis (A) increased 56% and photorespiration decreased36% in terminal mainstem leaves with CO₂ enrichment. Net photosynthesisand photorespiration were suppressed 30% and 41%, respectively,by elevated O₃ during late reproductive growth in the ambientCO₂ treatment, but not in the elevated CO₂ treatment. The ratioof photorespiration to A at growth CO₂ was decreased 61% byelevated CO₂ There was no statistically significant effect ofelevated O₃ on the ratio of photorespiration to A. Activitiesof glycolate oxidase, hydroxypyruvate reductase and catalasewere decreased 10–25% by elevated CO₂ and by 46–66%by elevated O₃ at late reproductive growth. The treatments hadno significant effect on total amino acid or glycine levels,although serine concentration was lower in the elevated CO₂and O₃ treatments at several sampling dates. The inhibitoryeffects of elevated O₃ on photorespiration-related parameterswere generally commensurate with the O₃-induced decline in A.The results suggest that elevated CO₂ could promote productivityboth through increased photoassimilation and suppressed photorespiration. Key words: Photorespiration, CO₂-enrichment, ozone, climate change, air pollution     

Correlation between AS1 gene expression and seed protein contents in different soybean (Glycine max [L.] Merr.) cultivars

In higher plants, asparagine synthetase (AS) plays an important role in regulating the nitrogen sink-source relationship. We studied the expression of AS genes in five Chinese soybean cultivars exhibiting contrasting seed protein contents. We found that only the AS2 but not the AS1 gene was induced by dark treatment. On the other hand, the expression of AS1 in leaves (especially in trifoliate leaves of young seedlings) showed a positive correlation with seed protein contents in the soybean cultivars tested. Therefore, in spite of the fact that the principle transporting compounds in soybean plants for nitrogen acquired via symbiotic fixation are ureides, AS may still play an important role in the process of nitrogen assimilation.     

Genistein accumulation in soybean (Glycine max [L.] Merr.) root systems under suboptimal root zone temperatures

Genistein, as a plant-to-bacteria signal, plays an importantrole in the establishment of the soybean (Glycine max [L.] Merr.)-Bradyrhizobiumjaponicum nitrogen-fixing symbiosis. It is essential to thedevelopment of effective root nodules and responsible for inducingthe nod genes of B. japonicum. Because sub-optimal root zonetemperature (RZT) delays infection and early nodule development,and decreases plant nodule number, and genistein addition overcomessome of this, it is reasonable to hypothesize that suboptimalRZT disrupts the inter-organismal signal exchange by inhibitinggenistein synthesis. Four experiments were conducted to testthese hypotheses. The results of these studies indicated that:(1) when soybean plants were germinated and maintained at RZTsranging from 13 to 17C, root genistein concentration and contentper plant were lower than those of plants with roots maintainedat RZTs above 17C; (2) when plants were germinated at an optimalRZT (25 C) then transferred to RZTs below 17C, and acclimatedfor a few days, root genistein concentration and content perplant were higher than those of plants with roots maintainedeither at optimal RZT, or transferred to RZT above 17 C, althoughby the end of the experiment, the genistein concentration ofroot systems at below 17C RZT appeared to be declining to valuesbelow those of plants with above 17 C RZT; (3) the root genisteinconcentration increased before the onset of nitrogen fixationand decreased thereafter; and (4) part of the effect of RZTson genistein content per plant root system was from reductionsin genistein concentration at lower RZT5, and part was due todecreased plant root growth. Key words: Genistein, Glycine max, suboptimal temperature     

Molecular mapping of k2 Mdh1-n y20, an unstable chromosomal region in soybean [Glycine max (L.) Merr.]

In the soybean genome, a chromosomal region covering three tightly linked genes, k2, Mdh1-n, and y20, was found very unstable. It was suspected that the instability of the k2 Mdh1-n y20 chromosomal region was caused by a non-autonomous transposable element residing adjacent to or in this region. In this study, we located and mapped this region with simple sequence repeat (SSR) markers on the soybean integrated map using five mapping populations. The k2 Mdh1-n y20 chromosomal region was located on molecular linkage group H. The integrated map from five mapping populations consisted of 13 loci in the order Satt541, Satt469, Sat_122, Satt279, Satt253, Satt314, Mdh1-n,y20, k2, Satt302, Satt142, Satt181, and Satt434. The k2 Mdh1-n y20 chromosomal region was very close to Satt314, Satt253, and Satt279. The genetic distance between the Mdh1-n gene and Satt314 was less than 1 cM. The results of the mapping study were consistent with the results from previous studies that the Mdh1-n mutation in T261 (k2 Mdh1-n) and the Mdh1-n y20 mutation in T317 (Mdh1-n y20) were caused by deletions. In addition, another putative deletion was found in the genome of T261 which covered three SSR markers (Satt314, Satt253, and Satt279). This is a joint contribution of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa, Project No. 3769, and from the USDA, Agricultural Research Service, Corn Insects and Crop Genetics Research Unit, and supported by the Hatch Act and the State of Iowa. The mention of a trademark or proprietary product does not constitute a guarantee or warranty of the product by Iowa State University or the USDA, and the use of the name by Iowa State University or the USDA implies no approval of the product to the exclusion of others that may also be suitable.     

Changes in Soybean (Glycine max [L.] Merr.) Glycerolipids in Response to Water Stress

Soybean (Glycine max [L.] Merr.) plants with the first trifoliate leaf fully expanded were exposed to 4 and 8 days of water stress. Leaf water potentials dropped from −0.6 megapascal to −1.7 megapascals after 4 days of stress; then to −3.1 megapascals after 8 days without water. All of the plants recovered when rewatered. The effects of short-term drought stress on triacylglycerol, diacylglycerol, phospholipid, and galactolipid metabolism in the first trifoliate leaves was determined. Leaf triacylglycerol and diacylglycerol content increased 2-fold during the first 4 days of stress and returned to control levels 3 days after rewatering. The polar lipid fraction, which contained phospholipids and galactolipids, changed little during this time. The linolenic acid (18:3) content of the triacylglycerol and diacylglycerol increased 25% during stress and the polar lipid 18:3 content decreased 15%. The pattern of glycerolipid labeling, after applying [2-¹⁴C]acetate to intact leaves was altered by water stress. After 4 days of water stress the radioactivity of phosphatidic acid + phosphatidylinositol, phosphatidylcholine, triacylglycerol, and diacylglycerol increased between 4 and 9% (compared to control plans) while radioactivity of phosphatidylethanolamine, monogalactosyldiglyceride, and digalactosyldiglyceride decreased 2 to 11%. These data indicated that increased levels of triacylglycerol and diacylglycerol observed during water stress were attributed to de novo synthesis rather than breakdown or reutilization of existing glycerolipids and fatty acids.     

Nitrate Reduction by Roots of Soybean (Glycine max [L.] Merr.) Seedlings

Studies were conducted with 9 to 12 day-old soybean (Glycine max [L.] Merr. cv. Williams) seedlings to determine the contribution of roots to whole plant NO(3) (-) reduction. Using an in vivo -NO(3) (-) nitrate reductase (NR) assay (no exogenous NO(3) (-) added to incubation medium) developed for roots, the roots accounted for approximately 30% of whole plant nitrate reductase activity (NRA) of plants grown on 15 mm NO(3) (-).Nitrogen analyses of xylem exudate showed that 53 to 66% of the total-N was as reduced-N, depending on the time of day of exudate collection. These observations supported enzyme data that suggested roots were contributing significantly to whole plant NO(3) (-) reduction. In short-term feeding studies using (15)N-NO(3) (-) significant and increasing atom percent (15)N excess was found in the reduced-N fraction of xylem exudate at 1.5 and 3 hours after feeding, respectively, which verified that roots were capable of reducing NO(3) (-).Estimated reduced-N accumulation by plants based on in vivo -NO(3) (-) NR assays of all plant parts substantially over-estimated actual reduced-N accumulation by the plants. Thus, the in vivo NR assay cannot be used to accurately estimate reduced-N accumulation but still serves as a useful assay for relative differences in treatment conditions.