Wheat Cultivar-Specific Selection of 2,4-Diacetylphloroglucinol-Producing Fluorescent \u3ci\u3ePseudomonas\u3c/i\u3e Species from Resident Soil Populations

An emerging body of evidence indicates a role for plant genotype as a determinant of the species and genetic composition of the saprophytic microbial community resident to the rhizosphere. In this study, experiments were conducted to determine the capacity of five different wheat cultivars to enhance resident populations and support introduced strains of 2,4-diacetylphloroglucinol (2,4-DAPG)-producing fluorescent pseudomonads, a group of bacteria known to provide biological control of several soilborne diseases. When soils were cropped with three successive 28-day growth cycles of wheat, the 2,4- DAPG-producing strains were consistently recovered from the rhizosphere of the cultivar Lewjain, and commonly were present at populations higher than those recovered from other wheat cultivars. Based on restriction fragment length polymorphism and sequence analyses of phlD, a key gene involved in 2,4-DAPG production, two previously undefined phlD+ genotypes, referred to as genotypes PfZ and PfY, were discovered. Wheat cultivar Lewjain was the primary source of genotype PfY while cultivar Penawawa yielded the majority of genotype PfZ. Based on 16S rDNA sequence analysis, both new phlD genotypes were classified as P. fluorescens. Comparison of the rhizosphere competence of 2,4-DAPG-producing P. fluorescens Q2-87 (genotype B) and P. fluorescens LR3-A28 (genotype PfY) showed that both strains persisted at similar populations in the rhizosphere of all cultivars tested over a 30 day period when introduced as a seed inoculant. However, when strain LR3-A28 was applied as a soil inoculant, this strain was recovered at higher populations from the rhizosphere of wheat cultivar Lewjain than from the rhizospheres of two other cultivars. No cultivar effects were shown for strain Q2-87. Collectively, these results add further to evidence indicating a degree of specificity in interactions between plant cultivars and specific members of the saprophytic microbial community. Furthermore, as 2,4- DAPG-producing fluorescent Pseudomonas spp. have a central role in the spontaneous reduction in severity of take-all disease of wheat in response to continuous wheat monoculture, we postulate that the use of specific cultivars, such as Lewjain, which possess a superior capacity to enhance resident soil populations of these bacteria may have potential to reduce the length of the monoculture period required to induce natural suppressiveness of soils toward this disease

Wheat Cultivar-Specific Selection of 2,4-Diacetylphloroglucinol-Producing Fluorescent \u3ci\u3ePseudomonas\u3c/i\u3e Species from Resident Soil Populations

An emerging body of evidence indicates a role for plant genotype as a determinant of the species and genetic composition of the saprophytic microbial community resident to the rhizosphere. In this study, experiments were conducted to determine the capacity of five different wheat cultivars to enhance resident populations and support introduced strains of 2,4-diacetylphloroglucinol (2,4-DAPG)-producing fluorescent pseudomonads, a group of bacteria known to provide biological control of several soilborne diseases. When soils were cropped with three successive 28-day growth cycles of wheat, the 2,4- DAPG-producing strains were consistently recovered from the rhizosphere of the cultivar Lewjain, and commonly were present at populations higher than those recovered from other wheat cultivars. Based on restriction fragment length polymorphism and sequence analyses of phlD, a key gene involved in 2,4-DAPG production, two previously undefined phlD+ genotypes, referred to as genotypes PfZ and PfY, were discovered. Wheat cultivar Lewjain was the primary source of genotype PfY while cultivar Penawawa yielded the majority of genotype PfZ. Based on 16S rDNA sequence analysis, both new phlD genotypes were classified as P. fluorescens. Comparison of the rhizosphere competence of 2,4-DAPG-producing P. fluorescens Q2-87 (genotype B) and P. fluorescens LR3-A28 (genotype PfY) showed that both strains persisted at similar populations in the rhizosphere of all cultivars tested over a 30 day period when introduced as a seed inoculant. However, when strain LR3-A28 was applied as a soil inoculant, this strain was recovered at higher populations from the rhizosphere of wheat cultivar Lewjain than from the rhizospheres of two other cultivars. No cultivar effects were shown for strain Q2-87. Collectively, these results add further to evidence indicating a degree of specificity in interactions between plant cultivars and specific members of the saprophytic microbial community. Furthermore, as 2,4- DAPG-producing fluorescent Pseudomonas spp. have a central role in the spontaneous reduction in severity of take-all disease of wheat in response to continuous wheat monoculture, we postulate that the use of specific cultivars, such as Lewjain, which possess a superior capacity to enhance resident soil populations of these bacteria may have potential to reduce the length of the monoculture period required to induce natural suppressiveness of soils toward this disease

Registration of N614, A3N615, N616, and N617 Shattercane Genetic Stocks with Cytoplasmic or Nuclear Male Sterility and Juicy or Dry Midribs

Four shattercane [Sorghum bicolor subsp. drummondii (Nees ex Steud.) de Wet ex Davidse] genetic stocks—N614 (Reg. No. GS-652, PI 665684), A3N615 (Reg. No. GS-651, PI 665683), N616 (Reg. No. GS-653, PI 665685), and N617 (Reg. No. GS-654, PI 665686)—with A3 cytoplasmic male sterility or the nuclear male sterility gene ms3 containing either juicy (dd) or dry (DD) culms were developed jointly by the USDA-ARS; the Iowa Agricultural and Home Economics Experiment Station, College of Agriculture and Life Sciences, Iowa State University; and the Agricultural Research Division, Institute of Agriculture and Natural Resources, University of Nebraska. The stocks were released in July 2011. The source material for these genetic stocks was isolated from an archetypical shattercane population found near Lincoln, NE. Release of these genetic stocks makes available shattercane lines with both A3 cytoplasmic male sterility, and ms3 genetic (nuclear) male sterility to facilitate crossing. These genetic stocks also contain juicy (dd) or dry (DD) culms, a visible genetic marker to facilitate screening progeny resulting from crosses. The genetic stocks have immediate application for basic research involving gene flow from cultivated sorghum [Sorghum bicolor (L.) Moench] to shattercane and on the fitness of offspring resulting from such crosses

Field damage of sorghum (\u3ci\u3eSorghum bicolor\u3c/i\u3e) with reduced lignin levels by naturally occurring insect pests and pathogens

Sorghum (Sorghum bicolor (L.) Moench) brown midrib (bmr) mutant lines have reduced levels of lignin, which is a potentially useful trait for bioenergy production, but the effects of this trait on insect and plant pathogen interactions are unknown under field conditions. Field-grown bmr6, bmr12, and wild-type (WT) plants were examined for insect and disease damage. In most cases, observed frequency, population, or leaf area damage caused by insects or pathogens on bmr6 or bmr12 plants were not greater than those observed on WT plants in the field or laboratory assays. European corn borers [Ostrinia nubilalis (Hübner)(Lepidoptera: Pyralidae)] often caused lower amounts of leaf damage to bmr6 leaves compared to bmr12 and sometimes WT leaves in the field study. Leaf damage by corn earworms [Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae)] and fall armyworms [Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae)] in laboratory assays was often lower for bmr versus WT leaves. Incidence of disease lesions was significantly higher on bmr6 compared to WT plants for one of three samplings in 2011, but the opposite trend was observed overall in 2012 and no significant differences were noted in 2013. When corn earworms and fall armyworms were fed the excised pith, bmr6 and/or bmr12 pith caused significant morality to one or both insect species in all 3 years. Damage variability between the 3 years may have been due to hotter and drier than normal conditions in 2012. Thus, bmr lines of sorghum suitable for bioenergy production have potential for sustainable production in the field

Efficacy of Singular and Stacked \u3ci\u3ebrown midrib 6\u3c/i\u3e and \u3ci\u3e12\u3c/i\u3e in the Modification of Lignocellulose and Grain Chemistry

In sorghum, brown midrib (bmr) 6 and 12 impair the last two steps of monolignol synthesis. bmr genes were introduced into grain sorghum to improve the digestibility of lignocellulosic tissues for grazing or bioenergy uses following grain harvest. Near-isogenic grain sorghum hybrids (AWheatland X RTx430) were developed containing bmr6, bmr12, and the bmr6 bmr12 double mutant (stacked), and their impacts were assessed in a two-year field study. The bmr genes did not significantly impact grain or lignocellulosic tissue yield. Lignocellulosic tissue from bmr6, bmr12, and stacked hybrids had reduced lignin content and increased in vitro dry matter digestibility (IVDMD) compared to those of the wild type (WT). The lignin content of the stacked lignocellulosic tissue was further reduced compared to that of bmr6 or bmr12. Surprisingly, bmr12 modestly increased carbohydrates in lignocellulosic tissue, and bmr6 increased fiber and lignin content in grain. These data indicate that bmr6 and bmr12 have broader effects on plant composition than merely lignin content, which has promising implications for both livestock utilization and bioenergy conversion

Efficacy of Singular and Stacked \u3ci\u3ebrown midrib 6\u3c/i\u3e and \u3ci\u3e12\u3c/i\u3e in the Modification of Lignocellulose and Grain Chemistry

In sorghum, brown midrib (bmr) 6 and 12 impair the last two steps of monolignol synthesis. bmr genes were introduced into grain sorghum to improve the digestibility of lignocellulosic tissues for grazing or bioenergy uses following grain harvest. Near-isogenic grain sorghum hybrids (AWheatland X RTx430) were developed containing bmr6, bmr12, and the bmr6 bmr12 double mutant (stacked), and their impacts were assessed in a two-year field study. The bmr genes did not significantly impact grain or lignocellulosic tissue yield. Lignocellulosic tissue from bmr6, bmr12, and stacked hybrids had reduced lignin content and increased in vitro dry matter digestibility (IVDMD) compared to those of the wild type (WT). The lignin content of the stacked lignocellulosic tissue was further reduced compared to that of bmr6 or bmr12. Surprisingly, bmr12 modestly increased carbohydrates in lignocellulosic tissue, and bmr6 increased fiber and lignin content in grain. These data indicate that bmr6 and bmr12 have broader effects on plant composition than merely lignin content, which has promising implications for both livestock utilization and bioenergy conversion

Field damage of sorghum (\u3ci\u3eSorghum bicolor\u3c/i\u3e) with reduced lignin levels by naturally occurring insect pests and pathogens

Sorghum (Sorghum bicolor (L.) Moench) brown midrib (bmr) mutant lines have reduced levels of lignin, which is a potentially useful trait for bioenergy production, but the effects of this trait on insect and plant pathogen interactions are unknown under field conditions. Field-grown bmr6, bmr12, and wild-type (WT) plants were examined for insect and disease damage. In most cases, observed frequency, population, or leaf area damage caused by insects or pathogens on bmr6 or bmr12 plants were not greater than those observed on WT plants in the field or laboratory assays. European corn borers [Ostrinia nubilalis (Hübner)(Lepidoptera: Pyralidae)] often caused lower amounts of leaf damage to bmr6 leaves compared to bmr12 and sometimes WT leaves in the field study. Leaf damage by corn earworms [Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae)] and fall armyworms [Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae)] in laboratory assays was often lower for bmr versus WT leaves. Incidence of disease lesions was significantly higher on bmr6 compared to WT plants for one of three samplings in 2011, but the opposite trend was observed overall in 2012 and no significant differences were noted in 2013. When corn earworms and fall armyworms were fed the excised pith, bmr6 and/or bmr12 pith caused significant morality to one or both insect species in all 3 years. Damage variability between the 3 years may have been due to hotter and drier than normal conditions in 2012. Thus, bmr lines of sorghum suitable for bioenergy production have potential for sustainable production in the field

Expression of the tobacco β- 1,3-glucanase gene, PR-2d, following induction of SAR with \u3ci\u3ePeronospora tabacina\u3c/i\u3e

Systemic acquired resistance (SAR) is induced following inoculation of Peronospora tabacina sporangia into the stems of Nicotiana tabacum plants highly susceptible to the pathogen. Previous results have shown that accumulation of acidic P-1,3-glucanases (PR-2\u27s) following induction of SAR by P. tabacina may contribute to resistance to P. tabacina. We showed that up-regulation of the PR-2 gene, PR-2d, following stem inoculation with P. tabacina, is associated with SAR. Studies using plants transformed with GUS constructs containing the full length promoter from PR-2d or promoter deletions, provided evidence that a previously characterized regulatory element that is involved in response to salicylic acid (SA), may be involved in regulation of PR-2d following induction of SAR with P. tabacina. This work provides evidence that regulation of PR-2 genes during P. tabacina-induced SAR may be similar to regulation of these genes during infection of N-gene tobacco by TMV or following exogenous application of SA, and provides further support for the role of SA in regulation of genes during P. tabacina-induced SAR

Expression of the tobacco β- 1,3-glucanase gene, PR-2d, following induction of SAR with \u3ci\u3ePeronospora tabacina\u3c/i\u3e

Systemic acquired resistance (SAR) is induced following inoculation of Peronospora tabacina sporangia into the stems of Nicotiana tabacum plants highly susceptible to the pathogen. Previous results have shown that accumulation of acidic P-1,3-glucanases (PR-2\u27s) following induction of SAR by P. tabacina may contribute to resistance to P. tabacina. We showed that up-regulation of the PR-2 gene, PR-2d, following stem inoculation with P. tabacina, is associated with SAR. Studies using plants transformed with GUS constructs containing the full length promoter from PR-2d or promoter deletions, provided evidence that a previously characterized regulatory element that is involved in response to salicylic acid (SA), may be involved in regulation of PR-2d following induction of SAR with P. tabacina. This work provides evidence that regulation of PR-2 genes during P. tabacina-induced SAR may be similar to regulation of these genes during infection of N-gene tobacco by TMV or following exogenous application of SA, and provides further support for the role of SA in regulation of genes during P. tabacina-induced SAR

Deleterious mutations predicted in the sorghum (\u3ci\u3eSorghum bicolor\u3c/i\u3e) \u3ci\u3eMaturity\u3c/i\u3e (\u3ci\u3eMa\u3c/i\u3e) and \u3ci\u3eDwarf\u3c/i\u3e (\u3ci\u3eDw\u3c/i\u3e) genes from whole‑genome resequencing

In sorghum [Sorghum bicolor (L.) Moench] the Maturity (Ma1, Ma2, Ma3, Ma4, Ma5, Ma6) and Dwarf (Dw1, Dw2, Dw3, Dw4) loci, encode genes controlling flowering time and plant height, respectively, which are critical for designing sorghum ideotypes for a maturity timeframe and a harvest method. Publicly available whole-genome resequencing data from 860 sorghum accessions was analyzed in silico to identify genomic variants at 8 of these loci (Ma1, Ma2, Ma3, Ma5, Ma6, Dw1, Dw2, Dw3) to identify novel loss of function alleles and previously characterized ones in sorghum germplasm. From~ 33 million SNPs and~ 4.4 million InDels, 1445 gene variants were identified within these 8 genes then evaluated for predicted effect on the corresponding encoded proteins, which included newly identified mutations (4 nonsense, 15 frameshift, 28 missense). Likewise, most accessions analyzed contained predicted loss of function alleles (425 ma1, 22 ma2, 40 ma3, 74 ma5, 414 ma6, 289 dw1, 268 dw2 and 45 dw3) at multiple loci, but 146 and 463 accessions had no predicted ma or dw mutant alleles, respectively. The ma and dw alleles within these sorghum accessions represent a valuable source for manipulating flowering time and plant height to develop the full range of sorghum types: grain, sweet and forage/biomass