The resistance of peanut to soil-borne pathogens improved by rhizosphere probiotics under calcium treatment.

BACKGROUND: Peanut (Arachis hypogaea L.) is an important oil and economic crop. Calcium modulates plants in response to abiotic stresses and improves plant resistance to pathogens. Enrichment of beneficial microorganisms in the rhizosphere is associated with plant disease resistance and soil development. The purpose of this study was to analyze the differences in peanut rhizosphere microbial community structure between the calcium treatment and the control during two growth stages and to explain why calcium application could improve the resistance of peanuts to soil-borne pathogens. RESULTS: The 16S rDNA amplicon sequencing of rhizosphere microbiome showed that calcium application significantly enriched Serratia marcescens and other three dominant strains at the seedling stage. At the pod filling stage, ten dominant stains such as Sphingomonas changbaiensis and Novosphingobium panipatense were enriched by calcium. Serratia marcescens aseptic fermentation filtrate was mixed with PDA medium and inoculated with the main soil-borne pathogens in the seedling stage, which could inhibit the growth of Fusarium solani and Aspergillus flavus. The aseptic fermentation filtrate of Novosphingobium panipatense was mixed with PDA medium and inoculated with the main soil-borne pathogens in the pod filling stage, which could inhibit the growth of Sclerotium rolfsii and Leptosphaerulina arachidicola. CONCLUSIONS: Calcium application increases the resistance of peanuts to soil-borne pathogens by enriching them with specific dominant bacteria.

ZnCl2 treatment improves nutrient quality and Zn accumulation in peanut seeds and sprouts.

Peanut is a popular food due to its high nutrient content. The effects of ZnCl2 on peanut seed germination, fatty acid and sugar contents, vitamin biosynthesis, antioxidant content, and Zn assimilation were evaluated in this study. Treatment with ZnCl2 significantly improved the germination rate, enhanced reactive oxygen species production and reduced the content of total fatty acids in peanut seed and sprout. However, ZnCl2 treatment did not reduce total sugar or total protein relative to the control. Germination promoted the biosynthesis of phenolics and resveratrol and increased the antioxidant capacity, as evaluated by Fe3+ reducing power and 2,2-diphenyl-1-picrylhydrazyl radical scavenging ability, especially under Zn stress conditions. The vitamin content decreased in the following order among treatments: germinated seeds with ZnCl2 treatment > germinated seeds without ZnCl2 treatment > dormant seeds. Interestingly, Zn content was approximately five times higher in the germinated ZnCl2-treated seeds compared to in the untreated germinated seeds and the dormant seeds. The results of this study provide a new method for producing healthy foods with enhanced vitamin content and antioxidant capacity.

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.

Generation of peanut drought tolerant plants by pingyangmycin-mediated in vitro mutagenesis and hydroxyproline-resistance screening.

In order to enlarge the potential resources of drought-tolerant peanuts, we conducted in vitro mutagenesis with Pingyangmycin (PYM) as the mutagen as well as directed screening on a medium supplemented with Hydroxyproline (HYP). After being extracted from mature seeds (cv. Huayu 20), the embryonic leaflets were cultured on somatic embryogenesis-induction medium with 4 mg/L PYM and the generated embryos were successively transferred to a germination medium with 4 and then 8 mmol/L HYP to screen HYP-tolerant plantlets. After that, these plantlets were grafted and transplanted to the experimental field. In the next generation, all seeds were sown in the field, and phenotype variation and trait segregation can be observed in most of the offspring (M2 generation). The M3 generation individuals were subjected to drought stress at the seedling stages. The activities of SOD and POD were substantially increased in eight offspring of 11 HYP-tolerant, regenerated plants than in their mutagenic parents. To determine the correlation between mutant phenotypes and genomic modification, we carried out a comparison of the DNA polymorphisms between the mutagenic parents and 13 M3 generation individuals from different HYP-tolerant, regenerated plants with SSR primers. Results showed that most mutants and parent plants had signs of polymorphisms. Under drought stress, some M3 generation individuals of 10 original HYP-tolerant, regenerated plants produced more pods than the mutagenic parent; twenty individuals among them produced >60 g pods/plant. M4-generation seeds were tested for quality characteristics by Near Infrared Spectroscopy (NIS) and nine individuals with higher protein content (>30%) and 21 individuals with higher oil content (>58%) were screened. We concluded that the use of PYM-based in vitro mutagenesis in combination with directed screening with HYP is effective for the creation of potential drought-tolerant mutants of peanut.

Serine/threonine/tyrosine protein kinase phosphorylates oleosin, a regulator of lipid metabolic functions.

Plant oils are stored in oleosomes or oil bodies, which are surrounded by a monolayer of phospholipids embedded with oleosin proteins that stabilize the structure. Recently, a structural protein, Oleosin3 (OLE3), was shown to exhibit both monoacylglycerol acyltransferase and phospholipase A(2) activities. The regulation of these distinct dual activities in a single protein is unclear. Here, we report that a serine/threonine/tyrosine protein kinase phosphorylates oleosin. Using bimolecular fluorescence complementation analysis, we demonstrate that this kinase interacts with OLE3 and that the fluorescence was associated with chloroplasts. Oleosin-green fluorescent protein fusion protein was exclusively associated with the chloroplasts. Phosphorylated OLE3 exhibited reduced monoacylglycerol acyltransferase and increased phospholipase A(2) activities. Moreover, phosphatidylcholine and diacylglycerol activated oleosin phosphorylation, whereas lysophosphatidylcholine, oleic acid, and Ca(2+) inhibited phosphorylation. In addition, recombinant peanut (Arachis hypogaea) kinase was determined to predominantly phosphorylate serine residues, specifically serine-18 in OLE3. Phosphorylation levels of OLE3 during seed germination were determined to be higher than in developing peanut seeds. These findings provide direct evidence for the in vivo substrate selectivity of the dual-specificity kinase and demonstrate that the bifunctional activities of oleosin are regulated by phosphorylation.

Interspecific root interactions and rhizosphere effects on salt ions and nutrient uptake between mixed grown peanut/maize and peanut/barley in original saline-sodic-boron toxic soil.

Two glasshouse studies were conducted to investigate the effect of interspecific complementary and competitive root interactions and rhizosphere effects on the concentration and uptake of Na, Cl and B, and N, P, K, Ca, Mg, Fe, Zn and Mn nutrition of mixed cropped peanut with maize (Experiment I), and barley (Experiment II) grown in nutrient-poor saline-sodic and B toxic soil. Mixed cropped plants were grown in either higher density or lower density. The results of the experiment revealed that dry shoot weight decreased in peanut but increased in maize and barley with associated plant species compared to their monoculture. Shoot Na and Cl concentrations of peanut decreased significantly in both experiments, regardless of higher or lower density. The concentrations of Na also decreased in the shoots of mixed cropped maize and barley, but Cl concentrations increased slightly. The concentration of B significantly decreased in mixed cropping in all plant species regardless of higher or lower density. Rhizosphere chemistry was strongly and differentially modified by the roots of peanut, maize and barley, and mixed growing. There were significant correlations between the root-secreted acid phosphatases (S-APase), acid phosphatase in rhizosphere (RS-APase) and rhizosphere P concentration (RS-P) in the both experiments. The Fe-solubilizing activity (Fe-SA) and ferric reducing (FR) capacity of the roots were generally higher in mixed culture relative to their monoculture, which improved Fe, Zn and Mn nutrition of peanut. Further, there were also significant correlations among FR, Fe-SA and RS-Fe concentrations. Peanut facilitated P nutrition of maize and barley, while maize and barley improved K, Fe, Zn and Mn nutrition of peanut grown in nutrient-poor saline-sodic and B toxic soil.