Endophytic and rhizospheric bacteria associated with Paspalum atratum and its potential for plant growth promotion with different phosphate sources.

The genus Paspalum belongs to the family Poaceae and has several species that are native to Brazil. The Paspalum Germplasm Bank (GB) of the Brazilian Agricultural Research Corporation comprises approximately 450 accessions from 50 species. Among these accessions, Paspalum atratum (BGP 308) has economic potential for forage purposes. However, the endophytic and rhizospheric microbial communities within this accession and their ability to promote plant growth remain unknown. The present study aimed to isolate the endophytic and rhizospheric bacteria associated with P. atratum and to assess their potential for plant growth improvement, so-called plant growth-promoting bacteria (PGPB). For the in vitro tests, the ability of nitrogen-fixing bacteria (NFB), phosphate solubilization (PS) and indoleacetic acid (IAA) production were evaluated. A total of 116 endophytic and rhizosphere bacteria were obtained from the isolation. In the in vitro tests, 43 (37.00%) of these isolates showed positive NFB, PS, and IAA results. These isolates were identified by 16S rDNA sequencing. The phosphate solubilization index (PSI) ranged from 2 to 3.61, all 43 strains performed biological nitrogen fixation and the IAA production ranged from 12.85 to 431.41 µg ml-1. Eight of these 43 isolates were evaluated in vivo in a greenhouse using P. atratum caryopsis. The pots were filled with soil prepared with three different phosphate sources and one control without phosphate. After growth, the plants were submitted to morphological, bromatological and chemical determination. Data were analyzed using analysis of variance (ANOVA) and principal component analysis (PCA). In the in vivo test, treatments 105 (Pseudomonas sp.) and 458 (Pseudomonas sp.) were the most significant for the crystalline phosphate source, 109 (Bacillus sp.) for the sedimentary phosphate source and, as for the soluble phosphate source most treatments that received bacterial isolates had higher phosphorus content in the dry matter than the uninoculated soluble phosphate control. The 105FCR (crystalline phosphate + Pseudomonas sp.), 109FSE (sedimentary phosphate + Bacillus sp.), and 110 FSE (sedimentary phosphate + Enterobacter sp.) treatments showed the best results for plant growth promotion. This work made it possible to determine the bacterial community associated with P. atratum (BGP308) and to obtain new potential plant growth-promoting strains.

Prevalence of Delirium in End-of-Life Palliative Care Patients: An Observational Study.

OBJECTIVES: The aim of this study was to assess the prevalence of delirium, using the Assessment Test for Delirium and Cognitive Impairment (4AT) in end-of-life palliative care patients. SUBJECTS AND METHODS: This retrospective cross-sectional study was conducted on end-of-life patients in a hospice or at home. All patients were evaluated with the 4AT for the presence of delirium. RESULTS: Of the 461 patients analyzed, 76 (16.5%) were inpatients and 83.5% (385) outpatients. The median age was 79.5 (72-86) years, and 51.0% were female. According to the 4AT score, 126 patients (27.3%) had delirium (A4T ≥4) at admission, 28 (36.8%) were inpatients, and 98 (25.5%) outpatients. Around 33.8% of the cancer inpatients had delirium, while 20.6% of the cancer outpatients had delirium. The prevalence of delirium varied according to the setting, clinical condition, and life expectancy. In addition, 55.0% (11) actively dying inpatients, within 3 days, had delirium, and 56.7% (17) outpatients had delirium; while among those with life expectancy longer than 4 days, 30.4% (17) inpatients and 22.8% (81) outpatients were with delirium. CONCLUSIONS: Our study confirms that delirium is common in cancer and noncancer palliative care patients. Further research on delirium in end-of-life palliative care patients should consider the complexity of palliative care of this population as well as of the characteristics of the settings.

Transference of multiple resistance to peanut through the development of cross-compatible complex hybrids of wild Arachis.

Peanut (Arachis hypogaea L.) is a tetraploid species with an A and B genome, while the majority of wild Arachis species are diploid with distinct genomes. In pre-breeding programs, one way to introgress interesting wild genes into peanut is by producing amphidiploids. This study aimed at the hybridization between distinct amphidiploids and their characterization, to combine high crossability with peanut, observed in some amphidiploids, with high pest and disease resistances observed in others. These new hybrids were called complex hybrids. Four amphidiploids previously obtained were crossed at four different combinations, and the derived complex hybrids were crossed with four peanut cultivars. Morphological, reproductive, chromosome complement, molecular markers for hybrid identification, phytopatological, and entomological characterizations were performed on the complex hybrids. All cross combinations resulted in complex hybrids. One complete complement of each diploid progenitor was confirmed in each hybrid. Plants of six distinct hybrid combinations were obtained between the complex hybrids and peanut. Based on morphological characterization, differences among progenies from distinct cross combinations were observed. Complex hybrids were considered more resistant to all diseases and pests than peanut cultivars. The simultaneous introgression of genes from four wild Arachis species into peanut was possible through the development of complex hybrids.

Coexpression and Transcriptome analyses identify active Apomixis-related genes in Paspalum notatum leaves.

BACKGROUND: Paspalum notatum exhibits both sexual and apomictic cytotypes and, thus, is considered a good model for studies of apomixis because it facilitates comparative approaches. In this work, transcriptome sequencing was used to compare contrasting P. notatum cytotypes to identify differential expression patterns and candidate genes involved in the regulation of expression of this trait. RESULTS: We built a comprehensive transcriptome using leaf and inflorescence from apomictic tetraploids and sexual diploids/tetraploids and a coexpression network based on pairwise correlations between transcript expression profiles. We identified genes exclusively expressed in each cytotype and genes differentially expressed between pairs of cytotypes. Gene Ontology enrichment analyses were performed to better interpret the data. We de novo assembled 114,306 reference transcripts. In total, 536 candidate genes possibly associated with apomixis were detected through statistical analyses of the differential expression data, and several interacting genes potentially linked to the apomixis-controlling region, genes that have already been reported in the literature, and their neighbors were transcriptionally related in the coexpression network. CONCLUSIONS: Apomixis is a highly desirable trait in modern agriculture due to the maintenance of the characteristics of the mother plant in the progeny. The reference transcriptome, candidate genes and their coexpression network identified in this work represent rich resources for future grass breeding programs.

Resistance to thrips (Enneothrips flavens) in wild and amphidiploid Arachis species.

Thrips (Enneothrips flavens) is a pest that causes severe damage and yield losses to peanut crop if not properly controlled. The main control method currently used by farmers is bi-weekly application of insecticides during crop development, which, in addition to its toxicity, is very costly. Thus, new sources of resistance must be identified in order to reduce the use of insecticides and effectively manage the pest. This study aimed to evaluate the occurrence and symptoms of E. flavens infestations in 12 accessions of 10 wild species of Arachis and nine amphidiploids, as well as to compare their morphoagronomic characteristics to those of commercial cultivars. To this end, we conducted experiments during two summer seasons, using a randomized block design with four replications. We conducted evaluations of the severity of infestation, noting visual symptoms of E. flavens and morphological and reproductive characteristics of the Arachis plants. Results indicated that wild accessions V 7635 (A. vallsii), V 13250 (A. kempff-mercadoi), K 9484 (A. batizocoi), Wi 1118 (A. williamsii), V 14167 (A. duranensis) and V 13751 (A. magna) are the most promising for obtaining useful new amphidiploids. Among the amphidiploids, An 12 (A. batizocoi x A. kempff-mercadoi)4x, An 9 (A. gregoryi x A. stenosperma) 4x, and An 8 (A. magna x A. cardenasii)4x showed high level of resistance to E. flavens. The identified thrips resistant wild and amphidiploid Arachis species may be used in future breeding program to produce thrips resistant peanut cultivars.

Recombinants from the crosses between amphidiploid and cultivated peanut (Arachis hypogaea) for pest-resistance breeding programs.

Peanut is a major oilseed crop worldwide. In the Brazilian peanut production, silvering thrips and red necked peanut worm are the most threatening pests. Resistant varieties are considered an alternative to pest control. Many wild diploid Arachis species have shown resistance to these pests, and these can be used in peanut breeding by obtaining hybrid of A and B genomes and subsequent polyploidization with colchicine, resulting in an AABB amphidiploid. This amphidiploid can be crossed with cultivated peanut (AABB) to provide genes of interest to the cultivar. In this study, the sterile diploid hybrids from A. magna V 13751 and A. kempff-mercadoi V 13250 were treated with colchicine for polyploidization, and the amphidiploids were crossed with A. hypogaea cv. IAC OL 4 to initiate the introgression of the wild genes into the cultivated peanut. The confirmation of the hybridity of the progenies was obtained by: (1) reproductive characterization through viability of pollen, (2) molecular characterization using microsatellite markers and (3) morphological characterization using 61 morphological traits with principal component analysis. The diploid hybrid individual was polyploidized, generating the amphidiploid An 13 (A. magna V 13751 x A. kempff-mercadoi V 13250)4x. Four F1 hybrid plants were obtained from IAC OL 4 × An 13, and 51 F2 seeds were obtained from these F1 plants. Using reproductive, molecular and morphological characterizations, it was possible to distinguish hybrid plants from selfed plants. In the cross between A. hypogaea and the amphidiploid, as the two parents are polyploid, the hybrid progeny and selves had the viability of the pollen grains as high as the parents. This fact turns the use of reproductive characteristics impossible for discriminating, in this case, the hybrid individuals from selfing. The hybrids between A. hypogaea and An 13 will be used in breeding programs seeking pest resistance, being subjected to successive backcrosses until recovering all traits of interest of A. hypogaea, keeping the pest resistance.

First microsatellite markers for Paspalum plicatulum (Poaceae) characterization and cross-amplification in different Paspalum species of the Plicatula group.

BACKGROUND: Paspalum plicatulum is a perennial rhizomatous grass with natural diploid and polyploid cytotypes. It is a member of Plicatula, which has historically been recognized as a highly complex group containing species of ecological, ornamental and forage importance. The complex nature of the P. plicatulum genome makes it a challenging species for genetic research. This study aimed to develop and characterize microsatellite molecular markers in P. plicatulum and to evaluate their transferability to other Plicatula group species. FINDINGS: Microsatellite sequences were identified from three enriched libraries from P. plicatulum. Specific primers were designed, and 25 displayed polymorphism when screened across 48 polyploid Paspalum spp. genotypes. The number of bands per locus ranged from 2 to 17, with a mean of 8.65. Private bands for each species were identified; the highest number of private bands was observed for P. plicatulum in 52% of the loci analyzed. The mean polymorphism information content of all loci was 0.69, and the mean discriminatory power was 0.82. Microsatellite markers were satisfactorily cross-amplified for the eight tested Plicatula-group Paspalum species, with P. atratum exhibiting the highest transferability rate (89.86%). STRUCTURE and Discriminant Analysis of Principal Components separated accessions into three groups but did not reveal separation of the accessions according to species. CONCLUSIONS: This study describes the first microsatellite markers in P. plicatulum, which are polymorphic, efficient for the detection and quantification of genetic variation, and show high transferability into other species of the Plicatula group. This set of markers can be used in future genetic and molecular studies necessary for the proper development of conservation and breeding programs. Private bands within the markers can be used to assist in species identification.

Successful crosses between fungal-resistant wild species of Arachis (section Arachis) and Arachis hypogaea.

Peanut (Arachis hypogaea) is the fifth most produced oil crop worldwide. Besides lack of water, fungal diseases are the most limiting factors for the crop. Several species of Arachis are resistant to certain pests and diseases. This study aimed to successfully cross the A-genome with B-K-A genome wild species previously selected for fungal disease resistance, but that are still untested. We also aimed to polyplodize the amphihaploid chromosomes; cross the synthetic amphidiploids and A. hypogaea to introgress disease resistance genes into the cultivated peanut; and analyze pollen viability and morphological descriptors for all progenies and their parents. We selected 12 A-genome accessions as male parents and three B-genome species, one K-genome species, and one A-genome species as female parents. Of the 26 distinct cross combinations, 13 different interspecific AB-genome and three AA-genome hybrids were obtained. These sterile hybrids were polyploidized and five combinations produced tetraploid flowers. Next, 16 combinations were crossed between A. hypogaea and the synthetic amphidiploids, resulting in 11 different hybrid combinations. Our results confirm that it is possible to introgress resistance genes from wild species into the peanut using artificial hybridization, and that more species than previously reported can be used, thus enhancing the genetic variability in peanut genetic improvement programs.

Identification of Fungus Resistant Wild Accessions and Interspecific Hybrids of the Genus Arachis.

Peanut, Arachis hypogaea L., is a protein-rich species consumed worldwide. A key improvement to peanut culture involves the development of cultivars that resist fungal diseases such as rust, leaf spot and scab. Over three years, we evaluated fungal resistance under field conditions of 43 wild accessions and three interspecific hybrids of the genus Arachis, as well as six A. hypogaea genotypes. In the first year, we evaluated resistance to early and late leaf spot, rust and scab. In the second and third years, we evaluated the 18 wild species with the best resistance scores and control cultivar IAC Caiapó for resistance to leaf spot and rust. All wild accessions displayed greater resistance than A. hypogaea but differed in their degree of resistance, even within the same species. We found accessions with as good as or better resistance than A. cardenasii, including: A. stenosperma (V15076 and Sv 3712), A. kuhlmannii (V 6413), A. kempff-mercadoi (V 13250), A. hoehnei (KG 30006), and A. helodes (V 6325). Amphidiploids and hybrids of A. hypogaea behaved similarly to wild species. An additional four accessions deserve further evaluation: A. magna (V 13751 and KG 30097) and A. gregoryi (V 14767 and V 14957). Although they did not display as strong resistance as the accessions cited above, they belong to the B genome type that is crucial to resistance gene introgression and pyramidization in A. hypogaea.

Resistência à mancha preta e qualidade agronômica de plantas rc1f2 de cruzamentos do híbrido anfidiplóide (Arachis ipaensis x A. duranensis) com o amendoim cultivado (A. hypogaea) / Resistance to late leafspot and agronomic quality of bc1f2 plants from crosses between the amphyploids hibrid (arachis ipaensis x a. duranensis) and the cultivated peanut (a. hypogaea)

A resistência à mancha preta (Cercosporidium personatum) e a condição agronômica de plantas de amendoim foram avaliadas em populações RC1F2 obtidas de cruzamentos do anfidiplóide (Arachis ipaensis x Arachis duranensis) com o genitor recorrente, cultivar Runner IAC 886. O experimento foi conduzido em condições de campo, em Pindorama, SP., sob condições naturais de ocorrência da doença. A doença foi avaliada em cada planta, em duas épocas, usando uma escala de notas visuais de 1 a 9, considerando o número de lesões nas folhas e a desfolha causada pela doença. Foi realizada após a colheita, em 90 plantas tomadas ao acaso, a avaliação da produção de vagens e grãos/planta, o rendimento de grãos após descascamento, massa média de grãos, número de vagens/planta, número e porcentagem de vagens com dois grãos. Os resultados mostraram ampla variabilidade para resistência à doença. Dependendo do genitor hypogaea utilizado nos cruzamentos iniciais com o anfidiplóide, 4,2 a 7,7% de plantas portadoras de resistência próxima a do parental silvestre resistente (A. ipaensis) foram resgatadas em F2. Observaram-se diferenças significativas entre plantas RC1F2 em função da planta F1 da qual descendem, tanto para resistência à doença como para caracteres agronômicos, tendo em vista a condição segregante das plantas F4 utilizadas nos cruzamentos com o genitor recorrente, cultivar Runner IAC 886. As correlações negativas observadas entre resistência à doença e caracteres agronômicos como produção de vagens e grãos/planta e massa média de grãos sugerem a necessidade de novo ciclo de retrocruzamentos para quebrar eventuais ligações indesejáveis e gerar indivíduos resistentes mais próximos do padrão comercial.

Resistance to late leafspot (Cercosporidium personatum) and agronomic condition of peanut plants were assessed in segregating RC1F2 populations obtained from crosses between the amphidiploid (A. ipaensis x A. duranensis) and the recurrent parent, cultivar Runner IAC 886. The experiment was carried out at Pindorama, SP, under natural field conditions of the disease. Disease was evaluated at two dates during the growing cycle, in 1,500 plants, using a scale of visual notes ranging from 1 to 9, and taking into consideration the number of lesions in leaves and the degree of defoliation caused by the disease. The data obtained were used to estimate the AUDPC (Area Under the Disease Progress Curve). Agronomic evaluations were performed after harvesting, in 90 randomly taken plants. The following agronomic characters were evaluated: pod and kernel production per plant, shelling percentage, number of pods per plant, number and percentage of two-seed pods. The results showed wide variability for resistance to the disease. Depending on the hypogaea parent used in the initial crosses with the amphidiploids, 4,2 to 7,7% of plants with disease resistance close to the wild resistant parent (A. ipaensis) could be rescued in the F2 generation. Significant differences were observed between RC1F2 plants descendent from the same F1 plant, in either disease resistance or in agronomic traits, due to the segregant condition of the F4 plants used in the crosses to the recurrent parent, cultivar Runner IAC 886. Some of the RC1F1:2 plant families exhibited agronomic condition close to the cultivar IAC Caiapó, especially in production of pods and kernels, shelling percentage and average kernel weight. However, the large majority of the segregants in this generation still showed high percentages of one-seed pods. This condition, and the negative correlations observed between resistance to the disease and agronomic traits such as pod and kernel production per plant and average kernel weight suggest that a new backcrossing cycle is needed to break eventual undesirable linkages and generate resistant individuals closer to the commercial standards.

Occurrence of Enneothrips flavens moulton and Stegasta bosquella (Chambers) and its effects on agronomic traits of wild Arachis accessions / Ocorrência de Enneothrips flavens moulton e Stegasta bosquella (Chambers) e seus efeitos em caracteres agronômicos de acessos de espécies silvestres de arachis

Some wild species of the genus Arachis have demonstrated potential for improvement of peanuts. This work was performed to evaluate the occurrence and symptoms of Enneothrips flavens and Stegasta bosquella and its effects on agronomic traits of wild Arachis accessions. Nine accessions of wild Arachis species and a commercial A. hypogaea variety were studied in a split plot statistical scheme with a completely randomized block design and four replications. The main plots consisted of plants sprayed or not sprayed for insect control, while the subplots comprised the peanut accessions. Accessions GKP10017 (A. cardenasii) and V7639 (A. kuhlmannii) showed the lowest percentages of leaflets with E. flavens and S. bosquella. Accessions V9912, V7639 and V8979 (all three A. kuhlmannii) and V13250 (A. kempff-mercadoi) showed the lowest responses to insecticide application among the various plant traits evaluated. These accessions are of interest for further studies to identify mechanisms of resistance, to be used in breeding programs for resistance to these insects.

Algumas espécies silvestres do gênero Arachis têm demonstrado potencial para melhoramento do amendoim cultivado. Objetivou-se avaliar a ocorrência e os sintomas de Enneothrips flavens e Stegasta bosquella e seus efeitos nos caracteres agronômicos de acessos de Arachis spp. O delineamento estatístico utilizado foi o de blocos ao acaso, em esquema de parcelas subdivididas, sendo a aplicação ou não de inseticidas as parcelas e os acessos (dez materiais) as subparcelas, com quatro repetições. Os acessos GKP10017 (A. cardenasii) e V7639 (A. kuhlmannii) apresentaram as menores percentagens de folíolos com presença de E. flavens e S. bosquella e os acessos V9912, V7639 e V8979 (todos A. kuhlmannii) e V13250 (A. kempff-meradoi) se destacaram por apresentarem baixa resposta em relação à aplicação de inseticidas nos diferentes parâmetros avaliados. Esses acessos são de interesse para estudar os possíveis mecanismos de resistência às pragas e aproveitá-los em programas de melhoramento visando resistência.

Identification of candidate genome regions controlling disease resistance in Arachis.

BACKGROUND: Worldwide, diseases are important reducers of peanut (Arachis hypogaea) yield. Sources of resistance against many diseases are available in cultivated peanut genotypes, although often not in farmer preferred varieties. Wild species generally harbor greater levels of resistance and even apparent immunity, although the linkage of agronomically un-adapted wild alleles with wild disease resistance genes is inevitable. Marker-assisted selection has the potential to facilitate the combination of both cultivated and wild resistance loci with agronomically adapted alleles. However, in peanut there is an almost complete lack of knowledge of the regions of the Arachis genome that control disease resistance. RESULTS: In this work we identified candidate genome regions that control disease resistance. For this we placed candidate disease resistance genes and QTLs against late leaf spot disease on the genetic map of the A-genome of Arachis, which is based on microsatellite markers and legume anchor markers. These marker types are transferable within the genus Arachis and to other legumes respectively, enabling this map to be aligned to other Arachis maps and to maps of other legume crops including those with sequenced genomes. In total, 34 sequence-confirmed candidate disease resistance genes and five QTLs were mapped. CONCLUSION: Candidate genes and QTLs were distributed on all linkage groups except for the smallest, but the distribution was not even. Groupings of candidate genes and QTLs for late leaf spot resistance were apparent on the upper region of linkage group 4 and the lower region of linkage group 2, indicating that these regions are likely to control disease resistance.

Genetic mapping of wild introgressions into cultivated peanut: a way toward enlarging the genetic basis of a recent allotetraploid.

BACKGROUND: Peanut (Arachis hypogaea L.) is widely used as a food and cash crop around the world. It is considered to be an allotetraploid (2n = 4x = 40) originated from a single hybridization event between two wild diploids. The most probable hypothesis gave A. duranensis as the wild donor of the A genome and A. ipaënsis as the wild donor of the B genome. A low level of molecular polymorphism is found in cultivated germplasm and up to date few genetic linkage maps have been published. The utilization of wild germplasm in breeding programs has received little attention due to the reproductive barriers between wild and cultivated species and to the technical difficulties encountered in making large number of crosses. We report here the development of a SSR based genetic map and the analysis of genome-wide segment introgressions into the background of a cultivated variety through the utilization of a synthetic amphidiploid between A. duranensis and A. ipaënsis. RESULTS: Two hundred ninety eight (298) loci were mapped in 21 linkage groups (LGs), spanning a total map distance of 1843.7 cM with an average distance of 6.1 cM between adjacent markers. The level of polymorphism observed between the parent of the amphidiploid and the cultivated variety is consistent with A. duranensis and A. ipaënsis being the most probable donor of the A and B genomes respectively. The synteny analysis between the A and B genomes revealed an overall good collinearity of the homeologous LGs. The comparison with the diploid and tetraploid maps shed new light on the evolutionary forces that contributed to the divergence of the A and B genome species and raised the question of the classification of the B genome species. Structural modifications such as chromosomal segment inversions and a major translocation event prior to the tetraploidisation of the cultivated species were revealed. Marker assisted selection of BC1F1 and then BC2F1 lines carrying the desirable donor segment with the best possible return to the background of the cultivated variety provided a set of lines offering an optimal distribution of the wild introgressions. CONCLUSION: The genetic map developed, allowed the synteny analysis of the A and B genomes, the comparison with diploid and tetraploid maps and the analysis of the introgression segments from the wild synthetic into the background of a cultivated variety. The material we have produced in this study should facilitate the development of advanced backcross and CSSL breeding populations for the improvement of cultivated peanut.