An efficient protocol for in vitro regeneration of peanut (Arachis hypogaea L ) cultivar L14

The present work aims to establish an efficient protocol for in vitro regeneration of peanut (Arachis hypogaea) cultivar L14. The study showed that de-embryonated cotyledon was a suitable explant for shoot multiplication on MS medium containing 4 mg/L BAP. The highest number of shoots per explant obtained after 4 weeks of culture was up to 6.8 shoots. Shoots in vitro were able to produce a large number of approximately 11 roots on MS medium supplemented with 0.5 mg/L NAA. These results will be very useful in establishing an in vitro regeneration protocol for peanut cultivar L14 during gene transfer in the next studies to improve their disease resistance.

Characterization of AhLea-3 and its enhancement of salt tolerance in transgenic peanut plants

BACKGROUND: Late embryogenesis abundant (LEA) proteins were reported to be related to adversity stress and drought tolerance. Lea-3 from Arachis hypogaea L. (AhLea-3) was previously found to be related to salt tolerance according to the result of transcriptome profiling and digital gene expression analysis. So, AhLea-3 was cloned and the salt tolerance was validated by transgenic peanut plants. RESULTS: AhLea-3 was isolated from M34, a salt-resistant mutant of peanut, with its cDNA as the template. AhLea-3 contains one intron and two extrons, and the full-length cDNA sequence contains 303 bp. AhLea3 was ligated to pCAMBIA1301 to obtain the overexpression vector pCAMBIA1301-AhLea-3, which was then transferred into peanut variety Huayu23. The expression level of AhLea-3, as determined by qRTPCR analysis, was >10 times higher in transgenic than in non-transgenic plants. Five days after they were irrigated with 250 mM NaCl, the transgenic plants showed less severe leaf wilting, higher activities of antioxidant enzymes (superoxide dismutase, peroxidase, and catalase), and lower malonic dialdehyde content than non-transgenic plants. Relative to non-transgenic plants, the transgenic plants had a higher photosynthetic net rate, stomatal conductance, and transpiration rate, and a lower intercellular CO2 concentration after salt stress treatment (250 mM NaCl). CONCLUSIONS: These results indicate that overexpression of AhLea-3 increased the salt tolerance of transgenic peanut plants. AhLea-3 might become a useful gene resource for the variety breeding of salinity tolerance in peanut.

Morphological, biochemical and histological effects of aqueous extracts of peanut (Arachis hypogaea) on swiss mice in different diets

ABSTRACT Purpose: To evaluate the morphological, biochemical, and histological effects of aqueous extracts of peanut (skinless and added to 1% skin) in Swiss mice submitted to a high-fat diet. Methods: Forty male Swiss mice were divided into four groups (n=10 per group): GI) normocaloric diet; GII) high-fat diet; GIII) high-fat diet + 0.5 mL of peanut extract; GIV) high-fat diet + 0.5 mL of peanut extract + 1% peanut skin. The animals were weighed weekly and euthanized after 12 weeks for histopathological and biochemical analyses. The study was approved by the Animal Use Ethics Committee. Results: The animals in the GIV group had higher body weight when compared to the other ones. Increase in total cholesterol in GIII, increase in blood glucose in groups GII, GIII and GIV, decrease in serum low-density lipoprotein (LDL) concentration in groups GI and GIV and increase in serum concentration of C-reactive protein in GII were seen. The presence of vacuolar fat deposits was found in animal livers from GII. Conclusions: The extracts improved the plasma concentrations of animals that received a high-fat diet, including preventing morphological damage to liver tissue. These benefits were enhanced by the association of peanut shells with the extract.

Efficacy of Eosin Dye Removal by Peanut Shell Agrowaste Adsorbent

Abstract Peanut shell (PS) which is an excessive waste-product from agricultural processes, it can be recycled to a natural adsorbent for example it uses as removal dyes. Synthetic dye effluent without improperly discharged from industries to the river cause wastewater and damage to living organisms, especially, anionic dyes are difficult removed by conventional treatments such as biological, chemical, oxidation, and physical-filtration. However, an adsorption treatment is widely used for decolorization of dyes and give the best results for removal of various types of dissolved coloring materials. This research was used Eosin Y (EO) for the anionic model of dyeing wastewater and used PS for agrowaste adsorbent. The purpose of this study was investigated the efficiency adsorption of EO removal by PS. This efficiency adsorption was measured by different PS dosages, contact times, adsorbate concentration and equilibrium data. The results can be concluded that the PS had the efficiency adsorption of EO removal due to the equilibrium adsorption capacity (qe) and the highest dose of PS were balanced to adsorption of dye. The highest EO removal percentage was found in 87.7%, the qe was 0.351 mg g-1 and can adsorb from 10 mg L-1 to 1.23 mg L-1 in 25 g L-1 of PS dose at 30 minutes. In addition, the PS structure was found in multi-layer and many porous which is suitable for adsorbent. The morphological examination of PS was shown before and after adsorption that not changed. Therefore, PS might be an alternative choice for removal dye, and be used for the recycle adsorbent agrowaste as a commercial product for adding their values.

Identification of miRNAs linked to peanut nodule functional processes

microRNAs (miRNAs) are non-coding small RNAs that regulate gene expression at post-transcriptional level.Thousands of miRNAs have been identified in legumes, but studies about miRNAs linked to peanut nodulefunctionality are scarce. In this work we analyzed transcriptional changes in peanut nodules to identifymiRNAs involved in functional processes of these organs. We found 32 miRNAs precursors differentiallyexpressed in nodules compared with roots, and predicted the potential targets of their corresponding maturemiRNAs. Among them, 20 belong to 14 conserved miRNAs families and 12 are Arachis hypogaea-specificmiRNAs. Expression levels of 3 miRNAs (ahy-miR399, ahy-miR159 and ahy-miR3508) were confirmedexperimentally by qPCR. We also demonstrated that the expression of these miRNAs was not affected byinoculation of a biocontrol bacterium or a fungal pathogen. The catalogue of differentially expressed miRNAprecursors and the expression of the corresponding mature miRNA potential targets in the nodules of A.hypogaea obtained in this work is a database of strong candidates, including A. hypogaea-specific miRNAs, forthe regulation of the nodule functionality. The analysis of their role in this process will certainly lead to thecharacterization of essential regulators in these particular aeschynomenoid nodules.

Incidência de aflatoxinas B1, B2, G1 e G2 em amendoins / Incidence of aflatoxins B1, B2, G1 and G2 in peanuts

Micotoxinas são substâncias tóxicas produzidas por fungos e encontradas nos alimentos. As micotoxinas mais tóxicas são as aflatoxinas, produzidas, principalmente por Aspergillus flavus. Estudos realizados no país demonstraram alta incidência dessas micotoxinas em produtos de amendoim, que representa risco à saúde da população. O objetivo do estudo foi avaliar a incidência de aflatoxinas B1, B2, G1 e G2 em amostras de amendoins comercializados na região Nordeste do Estado de São Paulo nos períodos de 1994-2001 e 2016-2017. O método utilizado para analisar as amostras no primeiro período foi extração líquido-líquido e cromatografia em camada delgada e no segundo foi utilizando colunas de imunoafinidade, cromatografia líquida com derivatização pós- coluna e detector por fluorescência. No levantamento de 1994-2001 das 82 amostras, 39% tiveram contaminação de aflatoxinas variando de 11 a 1556 µg/kg com 37% das amostras contendo níveis maiores que 20 µg/kg, enquanto na pesquisa de 2016-17, das 56 amostras, 38% apresentaram contaminação destas toxinas variando de 0,09 a 60,40 µg/kg com 13% das amostras contendo níveis maiores que 20 µg/kg. Os resultados dos dois períodos estudados indicam que houve uma diminuição na incidência e nível das aflatoxinas estudadas, embora esta contaminação em amendoim permaneça um problema de saúde pública. (AU)

Mycotoxins are toxic compounds produced by fungi found in food. The most toxic mycotoxins are the aflatoxins produced mainly by Aspergillus flavus . Studies carried out in Brazil showed a high incidence of these mycotoxins in peanut products, a fact that represents public health problems. The aim of the study was to evaluate aflatoxins B1, B2, G1, and G2 in samples of peanuts sold in cities of the Northeast of the State of São Paulo in the period from 1994 to 2001 and from 2016 to 2017. The samples of the first period were analyzed using liquid-liquid extraction and thin-layer chromatography and the second using immunoaffinity columns, post-column derivative liquid chromatography and fluorescence detector. In the 1994-2001 survey, among 82 samples, 39% presented aflatoxins contamination ranging from 11 to 1556 µg/kg with 37% with levels greater than 20 µg/kg whereas, in the 2016-17 survey, 38% of the 56 samples presented contamination of aflatoxins ranging from 0.09 to 60.40 µg/kg and 7 samples 13% containing aflatoxins levels higher than 20 µg/kg. The results indicated there was a decrease in the incidence and level of aflatoxins, but the contamination of aflatoxins in peanuts remains a public health problem. (AU)

Incidência de aflatoxinas B1, B2, G1 e G2 em amendoins / Incidence of aflatoxins B1, B2, G1 and G2 in peanuts

Micotoxinas são substâncias tóxicas produzidas por fungos e encontradas nos alimentos. As micotoxinas mais tóxicas são as aflatoxinas, produzidas, principalmente por Aspergillus flavus. Estudos realizados no país demonstraram alta incidência dessas micotoxinas em produtos de amendoim, que representa risco à saúde da população. O objetivo do estudo foi avaliar a incidência de aflatoxinas B1, B2, G1 e G2 em amostras de amendoins comercializados na região Nordeste do Estado de São Paulo nos períodos de 1994-2001 e 2016-2017. O método utilizado para analisar as amostras no primeiro período foi extração líquido-líquido e cromatografia em camada delgada e no segundo foi utilizando colunas de imunoafinidade, cromatografia líquida com derivatização pós-coluna e detector por fluorescência. No levantamento de 1994-2001 das 82 amostras, 39% tiveram contaminação de aflatoxinas variando de 11 a 1556 μg/kg com 37% das amostras contendo níveis maiores que 20 μg/kg, enquanto na pesquisa de 2016-17, das 56 amostras, 38% apresentaram contaminação destas toxinas variando de 0,09 a 60,40 μg/kg com 13% das amostras contendo níveis maiores que 20 μg/kg. Os resultados dos dois períodos estudados indicam que houve uma diminuição na incidência e nível das aflatoxinas estudadas, embora esta contaminação em amendoim permaneça um problema de saúde pública.

Mycotoxins are toxic compounds produced by fungi found in food. The most toxic mycotoxins are the aflatoxins produced mainly by Aspergillus flavus. Studies carried out in Brazil showed a high incidence of these mycotoxins in peanut products, a fact that represents public health problems. The aim of the study was to evaluate aflatoxins B1, B2, G1, and G2 in samples of peanuts sold in cities of the Northeast of the State of São Paulo in the period from 1994 to 2001 and from 2016 to 2017. The samples of the first period were analyzed using liquid-liquid extraction and thin-layer chromatography and the second using immunoaffinity columns, post-column derivative liquid chromatography and fluorescence detector. In the 1994-2001 survey, among 82 samples, 39% presented aflatoxins contamination ranging from 11 to 1556 μg/kg with 37% with levels greater than 20 μg/kg whereas, in the 2016-17 survey, 38% of the 56 samples presented contamination of aflatoxins ranging from 0.09 to 60.40 μg/kg and 7 samples 13% containing aflatoxins levels higher than 20 μg/kg. The results indicated there was a decrease in the incidence and level of aflatoxins, but the contamination of aflatoxins in peanuts remains a public health problem.

Genome-wide development of polymorphic microsatellite markers and their application in peanut breeding program

BACKGROUND: Cultivated peanut (Arachis hypogaea. L) represents one of the most important oil crops in the world. Although much effort has been expended to characterize microsatellites or Simple Sequence Repeats (SSRs) in peanut, the quantity and quality of the markers in breeding applications remain limited. Here, genome-wide SSR characterization and marker development were performed using the recently assembled genome of the cultivar Tifrunner. RESULTS: In total, 512,900 microsatellites were identified from 2556.9-Mb genomic sequences. Based on the flanking sequences of the identified microsatellites, 7757 primer pairs (markers) were designed, and further evaluated in the assembled genomic sequences of the tetraploid Arachis cultivars, Tifrunner and Shitouqi, and the diploid ancestral species, A. duranensis and A. ipaensis. In silico PCR analysis showed that the SSR markers had high amplification efficiency and polymorphism in four Arachis genotypes. Notably, nearly 60% of these markers were single-locus SSRs in tetraploid Arachis species, indicating they are more specific in distinguishing the alleles of the A and B sub-genomes of peanut. In addition, two markers closely related with purple testa color and 27 markers near to FAD2 genes were identified, which could be used for breeding varieties with purple testa and high-oleic acid content, respectively. Moreover, the potential application of these SSR markers in tracking introgressions from Arachis wild relatives was discussed. CONCLUSIONS: This study reported the development of genomic SSRs from assembled genomic sequences of the tetraploid Arachis Tifrunner, which will be useful for diversity analysis, genetic mapping and functional genomics studies in peanut

Effects of Repeated Administration of Extracts from Arachis hypogaea Hulls on Blood Parameters and Histological Organization of Heart, Liver and Kidneys of Rats

Place and Duration: Laboratory of Biochemical Pharmacodynamics, Faculty of Biosciences, University Félix Houphouët - Boigny, from March to June 2018. Methodology: Serial extraction was done using methanol and water as solvents. The study was evaluated by orally daily doses of extracts 300, 600 and 1200 mg/kg. The treatment for 28 days concerned seven groups of animals, the control group and six treated groups. Each group included six animals, 3 males and 3 females. Animals of control group didn’t have any treatment. Animals were observed for general behavioural and signs of abnormalities during the experiment. After all treatments, blood was collected for haematologi cal and biochemical analysis. Liver, kidney, and heart were removed, weighed for histological study. Results: The results showed that, there were not any significant (p ?0.05) changes in both the absolute and relative organ weights between the control and the test groups. Biochemical parameters were statistically equal in all groups. In addition, both extracts did not induce any significant effect on RBC and indices relating to it (HGB, PCV, MCV, MCH and MCHC) throughout the experimental period. But, there was a decrease ( 16.33±1.68) on WBC with methanolic extract compared to control ( 13.79±2.73 ). Histological exam ination of the liver, kidneys, and the heart showed normal organisation and structure of heart, kidneys and liver. Conclusion: It appears that the methanolic and aqueous extracts of hull of Arachis hypogaea did not produce any toxicity in oral subacute toxicity study. However, further studies are needed to confirm long term toxicities.

Methods of cultivation and technical coefficients of peanuts in the Bahian Recôncavo / Métodos de cultivo e coeficientes técnicos do amendoim no Recôncavo Baiano

The objective of this study was to collect information about peanut cultivation methods adopted by family farmers in the Bahian Recôncavo (region that surrounds the bay in the state of Bahia, Brazil), using a specific form and also direct information from farmers. An ethnobotanical survey was carried out in the mesoregion of the Bahian Recôncavo in the following municipalities: Conceição do Almeida, Cruz das Almas, Maragogipe, São Felipe, São Felix, Sapeaçú and Laje (Vale do Jequiriçá), where 60 local farmers answered a nonstructured questionnaire about peanut cultivation. The cultivation system was estimated based on technical coefficients related to peanut cultivation in the area evaluated. The peanut cultivation system is carried out by family farmers from the Bahian Recôncavo who employ outdated practices and do not use the required basic steps. The technical coefficients analyzed show that the cultivation of peanut is an unprofitable activity when considering the high cost of production in this region, even though productivity is higher than in the entire state as well as in the Northeast region. Farmers in the Bahian Recôncavo do not consider the actual production costs of peanuts since they use seeds from previous crops and do not account for the labor costs of family members. There is a lack of active and continuous technical assistance to family farmers, not only in relation to the cultivation system, but also the possibility of other destinations for peanuts produced in the region.(AU)

O objetivo deste estudo foi levantar informações sobre os métodos de cultivo da cultura do amendoim, adotados por agricultores familiares do Recôncavo Baiano, utilizando-se de um formulário específico e de informações diretas dos agricultores. Foi realizado um levantamento etnobotânico nos municípios da mesorregião do Recôncavo Baiano: Conceição do Almeida, Cruz das Almas, Maragogipe, São Felipe, São Félix, Sapeaçú e no município de Laje (Vale do Jequiriçá), onde 60 agricultores foram entrevistados, por meio de um questionário não estruturado, referente ao cultivo do amendoinzeiro no Recôncavo Baiano. A estimativa do sistema de cultivo foi realizada com base em coeficientes técnicos referentes ao cultivo do amendoinzeiro na área de estudo. O sistema de cultivo do amendoim é realizado por agricultores familiares do Recôncavo Baiano utilizando práticas ultrapassadas, destituídas das etapas básicas necessárias. Os coeficientes técnicos analisados demonstram que o cultivo do amendoinzeiro é uma atividade pouco lucrativa ao se levar em consideração o alto custo de produção nessa região, mesmo a produtividade sendo maior quando comparada com a do estado, assim como com a da região Nordeste. Os agricultores do Recôncavo Baiano não consideram os custos de produção real do amendoim, uma vez que utilizam sementes de cultivos anteriores e não contabilizam os custos de mão de obra dos familiares. Falta aos agricultores familiares assistência técnica atuante e continuada, não só com relação ao sistema de cultivo como também com a possibilidade de outras destinações para o amendoim produzido na região.(AU)

Physiological response of seeds of peanut genotypes to water deficiency / Resposta fisiológica de sementes de genótipos de amendoim submetidos a deficiência hídrica

Among the factors that may influence the germination process of seeds, water deficiency is considered the most important. The present study aims to evaluate the physiological response of peanut genotypes to water deficiency. The experiment was conducted in a completely randomized design with four replications in a 12 x 7 factorial scheme with 12 peanut genotypes and seven polyethylene glycol 6000 concentrations corresponding to osmotic potentials of 0.00; -0.10; -0.20; -0.40; -0.60; -0.80 and -1.00 MPa. The data were submitted to analysis of variance and Tukey's test at a 5% probability level and, when significant, to analysis of variance adjusted to a quadratic polynomial curve. The germination percentage, seed vigor evaluated by the speed germination index, mean germination time, root length and root dry matter of some genotypes were affected by the osmotic potentials. Among the genotypes, there is a differential response related to the water deficiency level. In relation to the physiological response of the seeds, the lineages 506, 599, and 870 and the cultivar IAC Tatu-ST have a higher tolerance of water deficit. The peanut genotypes begin to suffer the effects of water deficiency at the osmotic potential of -0.4 MPa

Entre os fatores externos que podem influenciar o processo germinativo das sementes, a deficiência hídrica é considerada o mais importante. Dessa forma, o objetivo do presente trabalho foi avaliar a resposta fisiológica de sementes de genótipos de amendoim submetidos à deficiência hídrica. O experimento foi conduzido em esquema fatorial 12 x 7, tendo como tratamentos sementes de 12 genótipos de amendoim submetidos a sete concentrações de polietilenoglicol 6000, nos potenciais osmóticos correspondentes a 0,00; -0,10; -0,20; -0,40; -0,60; -0,80; -1,00 MPa. Constatou-se que os genótipos são afetados pelos potenciais osmóticos e, entre os genótipos, há resposta diferenciada em relação ao nível de deficiência hídrica. Em relação à resposta fisiológica das sementes, as linhagens 506, 599, 870 e a cultivar IAC Tatu-ST possuem maiortolerância à deficiência hídrica. Todos os genótipos de amendoim começam a sofrerem os efeitos da deficiência hídrica a partir do potencial osmótico de -0,4 MPa para a maior parte das características avaliadas

A case report of mono-sensitization to peanut component Ara h6

Peanut is the most common food allergen in the US, affecting 1-2 % of the population and studies show that it is still on the rise.' Component testing has offered better insight into the likelihood of reactivity with exposure. Extensive literature shows Arachis hypogea (Ara h) 2 as being the most clinically significant component identified to correlate with reactivity with exposure to the peanut protein, however there is minimal research on the reactivity of Ara h6. This case report describes a patient with a clinical reaction to peanut as a toddler and subsequent positivity on annual skin testing with commercial peanut extract, likely confounded by positive birch with advancing age. Immuno CAP testing revealed a negative Ara h2 and positive Ara h6, describing mono-sensitization to Ara h 6 and high probability of clinical reactivity. The importance of this case is to raise awareness of other highly allergenic components in patients with peanut allergy.

Transcriptome and proteome analyses of resistant preharvest peanut seed coat in response to Aspergillus flavus infection

BACKGROUND: The infection of peanut (Arachis hypogaea L.) seed coat by the pathogenic fungus Aspergillus flavus has highly negative economic and health impacts. However, the molecular mechanism underlying such defense response remains poorly understood. This study aims to address this issue by profiling the transcriptomic and proteomic changes that occur during the infection of the resistant peanut cultivar J11 by A. flavus. RESULTS: Transcriptomic study led to the detection of 13,539 genes, among which 663 exhibited differential expression. Further functional analysis found the differentially expressed genes to encode a wide range of pathogenesis- and/or defense-related proteins such as transcription factors, pathogenesis-related proteins, and chitinases. Changes in the expression patterns of these genes might contribute to peanut resistance to A. flavus. On the other hand, the proteomic profiling showed that 314 of the 1382 detected protein candidates were aberrantly expressed as a result of A. flavus invasion. However, the correlation between the transcriptomic and proteomic data was poor. We further demonstrated by in vitro fungistasis tests that hevamine-A, which was enriched at both transcript and protein levels, could directly inhibit the growth of A. flavus. Conclusions: The results demonstrate the power of complementary transcriptomic and proteomic analyses in the study of pathogen defense and resistance in plants and the chitinase could play an important role in the defense response of peanut to A. flavus. The current study also constitutes the first step toward building an integrated omics data platform for the development of Aspergillus-resistant peanut cultivars

Breeding on a new peanut variety Yuhua91 with high oleic acid content / 生物工程学报

Yuhua91 is a new peanut variety with high oleic acid content bred by Qingdao Agricultural University. The crossing was conducted with Luhua11 as female parent and with Kainong1715, an F435-type variety with high oleic acid content as male parent. The real F1 hybrids were screened by sequencing on PCR amplification products, and those homozygotes with bb genotype in F2 populations were screened by the same sequencing method as above. The content of oleic and linoleic acid was measured on the kernels harvested from F2 single plants by near infrared ray method, and those kernels whose content of oleic was above 80%, oleic and linoleic acid ratio was above 10.0 were obtained and planted into a row, with pedigree method for subsequent selection breeding. Yuhua91 has some characters of small pod, light and obvious pod texture, 148.06 g per 100 pods, 63.31 g per 100 kernels, 75.15% shelling percentage, long elliptic seed kernel, pink seed coat, without crack, white endotesta. Its content of protein, oil, oleic acid, linoleic acid and palmitic acid was 26.57%, 52.72%, 80.40%, 2.50% and 5.57% respectively. Yuhua91 has other characters of strong seedlings, compact pod areas, and moderate resistance to leaf spot disease and bacterial wilt. Average pod yield is 215.79 kg per Mu, 15.27% higher than the control variety Huayu20. Average seed kernels yield is 157.33 kg per Mu, 21.64% higher than the control variety Huayu20. Yuhua 91 has been registered on department of agriculture in 2018, and the registration No. is GPD peanut (2018) 370210, fit for growing in Shandong Province.

Transcriptome profiling and digital gene expression analysis of genes associated with salinity resistance in peanut

Background: Soil salinity can significantly reduce crop production, but the molecular mechanism of salinity tolerance in peanut is poorly understood. A mutant (S1) with higher salinity resistance than its mutagenic parent HY22 (S3) was obtained. Transcriptome sequencing and digital gene expression (DGE) analysis were performed with leaves of S1 and S3 before and after plants were irrigated with 250 mM NaCl. Results: A total of 107,725 comprehensive transcripts were assembled into 67,738 unigenes using TIGR Gene Indices clustering tools (TGICL). All unigenes were searched against the euKaryotic Ortholog Groups (KOG), gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases, and these unigenes were assigned to 26 functional KOG categories, 56 GO terms, 32 KEGG groups, respectively. In total 112 differentially expressed genes (DEGs) between S1 and S3 after salinity stress were screened, among them, 86 were responsive to salinity stress in S1 and/or S3. These 86 DEGs included genes that encoded the following kinds of proteins that are known to be involved in resistance to salinity stress: late embryogenesis abundant proteins (LEAs), major intrinsic proteins (MIPs) or aquaporins, metallothioneins (MTs), lipid transfer protein (LTP), calcineurin B-like protein-interacting protein kinases (CIPKs), 9-cis-epoxycarotenoid dioxygenase (NCED) and oleosins, etc. Of these 86 DEGs, 18 could not be matched with known proteins. Conclusion: The results from this study will be useful for further research on the mechanism of salinity resistance and will provide a useful gene resource for the variety breeding of salinity resistance in peanut.

Analysis of the peanut transgenic offspring with depressing AhFAD2 gene / 生物工程学报

The delta-12 fatty acid desaturase (Δ¹² FAD or FAD2) is a key enzyme that catalyzes oleic acid to linoleic acid by dehydrogenation at Δ¹² position of fatty acid carbon chain. In peanut, reduction or loss of FAD2 activity could enhance the relative content of oleic acid in kernels, and improve the quality and oxidation stability of peanut kernels and products. RNA interference (RNAi) technology could lead to non-expression or down-regulated expression of AhFAD2 gene. We constructed two RNA interference expression vectors with the inverted repeat sequence of partial AhFAD2 gene, which were driven separately by cauliflower mosaic virus (CaMV) 35S promoter or soybean agglutinin lectin seed-specific promoter. Homozygous transgenic lines carrying the two constructs stably in genetics were developed by peanut genetic transformation. There were no significant differences between the transgenic lines and the control through investigating the main agronomic traits. We analyzed the transcriptional level expression of AhFAD2 gene in transgenic lines and the control by real-time fluorescence quantitative PCR (qRT-PCR). The results suggested that the target genes of transgenic lines were likely suppressed by RNA interference, but showed different transcriptional levels in different peanut transgenic lines. Compared with untransformed lines, the resulting down-regulation of AhFAD2 gene resulted in a 15.09% or 36.40% increase in oleic acid content in the seeds of transformed HY23 and FH1 lines respectively, and the content of linoleic acid decreased by 16.19% or 29.81%, correspondingly, the ratio of oleic acid and linoleic acid (O/L) improved by 38.02%, 98.10%. The oleic acid content had significant differences between the two transformation constructs, and also among different transgenic lines. Moreover, the inhibition effect of RNAi was more obvious in the transgenic lines with FH1 as the receptor, and with transformation structure driven by seed specific promoter. The suppressed expression of AhFAD2 gene enabled the development of peanut fatty acid, which indicated that RNA interference would be a reliable technique for the genetic modification of peanut seed quality and the potential for improvement of other traits as well.

Ethnobotany of the peanut in recôncavo da Bahia, Brazil / Etnobotânica do amendoim no recôncavo da Bahia, Brasil

The aim of this study was to perform an ethnobotanical survey in some regions of Recôncavo da Bahia, Brazil, to investigate the profile of farmers and the local knowledge about Arachis hypogaea L. (peanut). We sampled 60 local peanut farmers from rural areas in the municipalities Conceição do Almeida, Cruz das Almas, Maragogipe, São Felipe, São Félix, Sapeaçú and Lage. One producer per residence was interviewed and a semi-structured interview model was used. During the interviews, seeds stored by the producers were collected for future analysis. The frequency of the answers was calculated using SAS (Statistical analysis system) software. Using an ethnobotanical approach, we were able to show that peanut cultivation is part of the Recôncavo da Bahia culture and that farmers in the area have their own knowledge about the use and management of this crop. This knowledge arose from information passed down over the years from previous generations. Similar farming techniques were used in all of the studied municipalities.

Objetivou-se realizar um levantamento etnobotânico em alguns municípios do Recôncavo da Bahia visando uma investigação sobre o perfil dos agricultores, além da caracterização do conhecimento local sobre a espécie. Foram feitas visitas a 60 produtores residentes em comunidades da zona rural dos municípios de Conceição do Almeida, Cruz das Almas, Maragogipe, São Felipe, São Félix, Sapeaçú e Lage, que têm contato direto com a cultura do amendoim, sendo entrevistada uma pessoa por domicílio, e que cultiva o amendoim. Utilizou-se um modelo de entrevista semiestruturada. No momento das entrevistas, foram realizadas as coletas de sementes armazenadas pelos produtores para serem testadas em trabalhos futuros. Foi calculada a frequência percentual de acordo com as respostas dos agricultores com auxílio do programa estatístico SAS - Statistical Analysis System. Por meio do levantamento etnobotânico, fica claro que o amendoim faz parte da cultura do Recôncavo da Bahia e que os agricultores detêm conhecimentos próprios sobre as formas de uso e manejo da cultura, oriundos de informações repassadas ao longo dos anos, vindas de gerações anteriores, levando-os a utilizarem técnicas de cultivo parecidas em todos os municípios que contribuíram com a pesquisa.

Development and application of KASP marker for high throughput detection of AhFAD2 mutation in peanut

Background: Cultivated peanut (Arachis hypogaea L.) is a major oilseed crop worldwide. Fatty acid composition of peanut oil may affect the flavor and shelf life of the resulting food products. Oleic acid and linoleic acid are the major fatty acids of peanut oil. The conversion from oleic acid to linoleic acid is controlled by theΔ12 fatty acid desaturase (FAD) encoded byAhFAD2AandAhFAD2B, two homoeologous genes from A and B subgenomes, respectively. One nucleotide substitution (G:C→A:T) ofAhFAD2Aand an "A" insertion ofAhFAD2Bresulted in high-oleic acid phenotype. Detection ofAhFAD2mutation had been achieved by cleaved amplified polymorphic sequence (CAPS), real-time polymerase chain reaction (qRT-PCR) and allele-specific PCR (AS-PCR). However, a low cost, high throughput and high specific method is still required to detectAhFAD2genotype of large number of seeds. Kompetitive allele specific PCR (KASP) can detect both alleles in a single reaction. The aim of this work is to develop KASP for detectionAhFAD2genotype of large number of breeding materials. Results: Here, we developed a KASP method to detect the genotypes of progenies between high oleic acid peanut and common peanut. Validation was carried out by CAPS analysis. The results from KASP assay and CAPS analysis were consistent. The genotype of 18 out of 179 BC4F2seeds was aabb. Conclusions: Due to high accuracy, time saving, high throughput feature and low cost, KASP is more suitable fordeterminingAhFAD2genotype than other methods.

Antioxidant activity, total phenolic, and resveratrol content in five cultivars of peanut sprouts

Objective To investigate the change in total phenolic compounds, antioxidant activity, and resveratrol content of five different germinated peanut cultivars. Methods The germinated sprouts of five peanut cultivars (Kalasin1, Kalasin2, Konkaen, Konkaen4, and Tainan9) were extracted with 80% ethanol and collected as crude extract. The antioxidant capacities were determined with 2,2-diphenyl-1-picrylhydrazyl and ferric ion reducing antioxidant power method. The total phenolic compound was measured using the Folin–Ciocalteau assay. The qualification and quantification of resveratrol was performed by high performance liquid chromatography method. Results Among the five cultivars, a three-day germination of Kalasin1 showed the highest phenolic content [(40.67 ± 2.62) μg gallic acid/g dry weight], expressed the highest 2,2-diphenyl-1-picrylhydrazyl antioxidant value [(80.51 ± 1.47) mmol/L Trolox/g dry weight], and ferric ion reducing antioxidant power antioxidant value [(171.33 ± 8.59) mmol/L ascorbic acid/g dry weight]. However, the high performance liquid chromatography result of Kalasin2 significantly increased to the highest resveratrol content of (6.44 ± 1.26) μg/g dry weight on the second day of germination. Conclusions The variation of phytochemical content in the peanut sprout is due to the effect of the peanut cultivar and the germination period.

Antioxidant activity, total phenolic, and resveratrol content in five cultivars of peanut sprouts

Objective:To investigate the change in total phenolic compounds,antioxidant activity,and resveratrol content of five different germinated peanut cultivars.Methods:The germinated sprouts of five peanut cultivars (Kalasinl,Kalasin2,Konkaen,Konkaen4,and Tainan9) were extracted with 80％ ethanol and collected as crude extract.The antioxidant capacities were determined with 2,2-diphenyl-l-picrylhydrazyl and ferric ion reducing antioxidant power method.The total phenolic compound was measured using the Folin-Ciocalteau assay.The qualification and quantification of resveratrol was performed by high performance liquid chromatography method.Results:Among the five cultivars,a three-day germination of Kalasinl showed the highest phenolic content [(40.67 ± 2.62) μg gallic acid/g dry weight],expressed the highest 2,2-diphenyl-l-picrylhydrazyl antioxidant value [(80.51 ± 1.47) mmol/L Trolox/g dry weight],and ferric ion reducing antioxidant power antioxidant value [(171.33 ± 8.59)mmol/L ascorbic acid/g dry weight].However,the high performance liquid chromatography result of Kalasin2 significantly increased to the highest resveratrol content of (6.44 ± 1.26) μg/g dry weight on the second day of germination.Conclusions:The variation of phytochemical content in the peanut sprout is due to the effect of the peanut cultivar and the germination period.