Comparison of the Δ^(12) fatty acid desaturase gene between high-oleic and normal-oleic peanut genotypes

△^12 fatty acid desaturase gene has been targeted as a logical candidate controlling the high oleate trait in peanut seeds. By RT-PCR method, the full-length cDNAs of △^12 fatty acid desaturase gene were isolated from peanut （Arachis hypogaea L.） genotypes with normal and high ratio of oleic to linoleic acid, which were designated AhFAD2B and AhFAD2B＇, respectively. Sequence alignment of their coding regions revealed that an extra A was inserted at the position ＋442 bp of AhFAD2B＇ sequence of high oleic acid genotypes, which resulted in the shift of open reading frame and a truncated protein AhFAD2B＇, with the loss of one histidine box involved in metal ion complex required for the reduction of oxygen. Analysis of transcript level showed that the expression of △^12 fatty acid desaturase gene in high oleic acid genotype was slightly lower than that in normal genotype. The enzyme activity experiment of yeast （Saccharomyces cerevisiae） cell transformed with AhFAD2B or AhFAD2B＇ proved that only AhFAD2B gene product showed significant △^12 fatty acid desaturase activity, but AhFAD2B＇ gene product did not. These results suggested that the change of AhFAD2B＇ gene sequence resulted in lower activity or deactivation of △^12 fatty acid desaturase in high oleic acid genotype.

Protective effect of high-oleic acid peanut oil and extra-virgin olive oil in rats with diet-induced metabolic syndrome by regulating branched-chain amino acids metabolism

High-oleic acid peanut oil(HOPO)and extra-virgin olive oil(EVOO)have been reported previously to have an attenuating effect on metabolic syndrome(MS).This study aimed to evaluate the metabolic effect of HOPO and EVOO supplementation in attenuating MS and the role of gut microbiota in regulating the metabolic profile.Sprague-Dawley rats were continuously fed with a normal diet,high-fructose and high-fat(HFHF)diet,HFHF diet containing HOPO,or a HFHF diet containing EVOO for 12 weeks.The metabolomics profiles of feces and serum samples were compared using untargeted metabolomics based on UPLC-Q/TOF-MS.Partial Least Squares Discriminant Analysis(PLS-DA)was used to identify the potential fecal and serum biomarkers from different groups.Correlation between gut microbiota and biomarkers was assessed,and pathway analysis of serum biomarkers was conducted.Differences in metabolic patterns in feces and serum were observed among different groups.There were 8 and 12 potential biomarkers in feces and 15 and 6 potential biomarkers in serum of HOPO group and EVOO group,respectively,suggesting that HOPO and EVOO supplementation mainly altered amino acids,peptides,and their analogs in feces and serum.The branched-chain amino acids(BCAAs)biosynthesis pathway was identified as a major pathway regulated by HOPO or EVOO.This study suggests that HOPO and EVOO supplementation ameliorate diet-induced MS,mainly via modulation of the BCAAs biosynthesis pathway.

Evaluation of allergenic protein profiles in three Chinese high-oleic acid peanut cultivars using NanoLC-Orbitrap mass spectrometry

High oleic-acid peanuts are known for their pre-longed shelf-life and health benefit due to high content of oleic fatty acid.However,the allergenicity and allergenic protein profiles in Chinese high-oleic peanuts have yet to be studied.For this purpose,an Orbitrap Fusion mass spectrometry(MS)-based method that is feasible for identification of putative allergenic protein as well as semi-quantitation of five major allergen protein in three different Chinese high-oleic peanut cultivars(JH 13,JH 16 and JH 18)have been reported.Results show that three Chinese high-oleic acid peanut cultivars selected all contained highly allergenic proteins Ara h1,Ara h 2,Ara h 3 and Ara h 6.The allergenic protein profiles of Chinese high-oleic acid peanut cultivars were very similar to that of conventional peanuts,but the allergenic protein subunits varied greatly among higholeic peanuts.Additionally,a comprehensive peptide-filtering pipeline had been developed for identification of potential peptide markers in peanut allergen proteins.Through the peptide-filtering pipeline,three novel peptide markers,IVQIEAKPNTLVLPK,SSNPDIYNPQAGSLR and AQSENYEYLAFK surrogate to Ara h 1,Ara h 3 with high abundance,good MS response and highly reliability were identified,which can be used as candidate peptide markers for the detection of peanut allergens in different food matrices.

Improving chemical and sensory quality of high-oleic peanut by application of foliar fertilizer

High-oleic peanut is favored by consumers because its a longer shelf life and benefits for cardiovascular health.Up to the high-oleic standard and with good taste is essential for peanut products in food processing.The aim of present study was to explore the effect of foliar fertilizers on chemical and eating quality of high-oleic peanuts.Three high-oleic peanut cultivars,namely Huayu 09C2,Huayu 9621,and Huayu 962,were used as materials.Huayu 09C2 and Huayu 9621 were planted in Fuxin,Liaoning,foliar sprayed with Fosforil at 2250 mL/ha,and Huayu 962 was sown in Jvnan,Shandong,foliar sprayed with Foliplus at 1500/3000 mL/ha.The chemical and sensory quality of peanut harvested from the treatment plots and controls was evaluated.Results showed that,Huayu 09C2 and Huayu 9621,foliar spraying with Fosforil in Fuxin,Huayu 962,foliar spraying with Foliplus in Jvnan,all still maintained high oleate content levels,and the Fosforil treatment was effective in raising the intensity of sweetness of roasted kernels.This study demonstrated the feasibility of improving chemical and sensory quality of high-oleic peanut through foliar fertilizer.

Comparison of anthocyanin and fatty acid in black and ordinary peanuts

In this study, anthocyanin and fatty acid compositions between black and ordinary peanuts were compared. Results showed that black peanuts contained 93.90±18.36 μg/g of cyanidin, which was significantly higher than that of ordinary peanuts. In addition, although both of black and ordinary peanuts shared some specific fatty acids, 4 fatty acids in black peanuts were significantly different from those in ordinary peanuts, which could be used as markers to distinguish black peanut from ordinary ones. This study might serve as a reference for authenticity identification and further nutritional researches on black peanut.

Esterification of cyclohexene with formic acid over a peanut shell-derived carbon solid acid catalyst

A carbon solid acid catalyst was prepared by the sulfonation of partially carbonized peanut shell with concentrated H2SO4. The structure and acidity of the catalyst were characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, X‐ray diffraction, thermogravimetric analysis, X‐ray photoelectron spectroscopy, and elemental analysis, which showed that it was an amorphous carbon material composed of aromatic carbon sheets in random orientations. Sulfonic acid groups were present on the surface at a density of 0.81 mmol/g. The carbon solid acid catalyst showed better performance than HZSM‐5 for the esterification of cyclohexene with formic acid. At a3：1 molar ratio of formic acid to cyclohexene, catalyst loading of 0.07 g/mL of cyclohexene, and reaction time of 1 h at 413 K, the cyclohexene conversion was 88.4% with 97.3% selectivity to cyclohexyl formate. The carbon solid acid catalyst showed better reusability than HZSM‐5 because its large pores were minimally affected by the accumulation of oligomerized cyclohexene, which deactivated HZSM‐5. The activity of the carbon solid acid catalyst decreased somewhat in the first two recycles due to the leaching of polycyclic aromatic hydrocarbon containing –SO3H groups and then it remained constant in the following reuse.

Inheritance of Fat and Fatty Acid Compositions in Peanut (Arachis hypogaea L.)

[Objective] The aim was to explore the inheritance of fat and fatty acid compositions in peanut.[Method] Genetic and correlation analyses were performed using a mixed genetic model of major gene plus polygene with 215 F9 RILs and their parents ZHENG 8903 and YUHUA No.4.[Result] The optimal genetic model of the fat content was two linked complementary major gene plus polygene inheritance model.The heritability of major genes was 22.88%.Oleic,linoleic,and palmitic acid content were controlled by two major genes and the heritability was 75.11%,75.00% and 60.95%,respectively.Stearic and arachidic acid content were controlled by two major genes plus polygenes,the heritability of major genes was 26.43% and 33.17%,respectively.O/L ratio and behenic acid content were controlled by three major genes,the heritability was 70.65% and 30.70%,respectively.[Conclusion] The result in this study was that accumulating polygenes should be emphasized in the improvement of fat content,while utilization of major genes is enhanced in improvement of oleic and linoleic acids.

Construction of a High-efficient Expression Vector of Δ^(12) Fatty Acid Desaturase in Peanut and Its Prokaryotical Expression

A full-length sequence coding for △^12 fatty acid desaturase gene from peanut（Arachis hypogaea L.）was cloned into the expression vector, pRSETB, to generate recombinant plasmid pRSET/HO-A, which was subsequently transformed into expression Escherichia. coli BL21（DE3）pLysS. The △^12 fatty acid desaturase was highly expressed in E. coli BL21（DE3）pLysS in the presence of isopropyl-D-thiogalactopyranoside （IPTG）. The fusion protein was purified and used to form a reaction system in vitro by adding oleic acid as substrate and incubating it at 20℃ for 6 h. Total fatty acids was extracted and methlesterified and then analyzed with gas chromatography. A novel peak corresponding to linoleic acid methyl ester standards was detected with the same retention time. GC-MS （gas chromatogram and gas chromatogram-mass spectrometry） analysis showed that the novel peak was linoleic acid methyl ester. These results exhibited △^12 fatty acid desaturase activity, which could convert oleic acid to linoleic acid specifically.

A comparison of methods used to determine the oleic/linoleic acid ratio in cultivated peanut (Arachis hypogaea L.)

There is an increasing demand in the peanut industry for high oleic peanuts and also for the incorporation of the high oleate trait into newly released varieties. Early generation screening of breeding lines for high oleic acid content greatly increases the efficiency of developing new peanut varieties. The objective of this study was to compare the accuracy of methods used to classify individual peanut seed as high oleic or not high oleic. Results from capillary electrophoresis (CE), two variations of near-infrared reflectance spectroscopy (NIRS), and real-time polymerase chain reaction (RT-PCR) genotyping, were compared with the traditionally accepted reference standard results from gas chromatography (GC). Three hundred and seventy-four (374) seeds, spanning twenty-three (23) genotypes and all four peanut market-types (runner, Spanish, Valencia and Virginia), were individually tested by each method. Percent accuracy levels for rating individual seed as high oleic (H) ranged from 97.4% (NIRS) to 99.5% (CE). All of the methods examined in this study carry only a minor risk for miss-classification (loss of material) and are suitable for use by peanut breeding programs in early generation breeding line screening.

Biochemical Composition and Disease Resistance in Newly Synthesized Amphidiploid and Autotetraploid Peanuts

Genetic diversity in peanut (Arachishypogaea L.) is narrow due to its evolution and domestication processes. Amphidiploids and autotetraploids (newly synthesized tetraploids) were created to broaden its genetic base. Molecular analysis has shown that the newly synthesized tetraploids had broader genetic base;and were genetically divergent when compared to cultivated peanut. Nutritional composition relative to oil, fatty acid composition, O/L ratio, protein, iodine value and presence of plant proteinase inhibitors such as trypsin and chymotrypsin inhibitors were studied in the synthesized tetraploids. Some of the newly synthesized tetraploids had higher amounts of proteinase inhibitors. Evaluation of newly synthesized tetraploids revealed several lines resistant to late leaf spot (LLS) and peanut bud necrosis disease (PBND).

Quantitative Trait Loci(QTL)Mapping and Marker Analysis of Fatty Acids in Peanut

Peanut,with high oil content,has been a major oil and food crop globally.The compositions of the fatty acids are the common factors in determining the oil quality.In the present study,an F2 segregated population with 140 individuals derived from the cross of Weihua8(a cultivar)and 12L49(a line with high oleic acid concentration)was used to construct a genetic map and conduct QTL mapping analysis.A total of 103 polymorphic SSR primers were utilized for genotyping the RILs and finally generating the SSR loci.Within the 103 SSR loci,a genetic linkage map,covering a total length of 3592.35 cM of the whole peanut genome,was constructed.Based on the genetic map,sixteen QTLs located on nine linkage groups related to peanut fatty acids were finally identified.Among them,four QTLs were detected associated with various traits simultaneously,which showed genetic stability in relation to fatty acids of peanut.Except for the QTLs for oleic acid,linoleic acid,and linolenic acid,three novel QTLs for arachidic acid and behenic acid were also detected.These QTLs might be helpful for further fine mapping analysis and marker-assisted selection of fatty acids in peanut.

Biochemical and microbial properties of rhizospheres under maize/ peanut intercropping

Maize/peanut intercropping system shows the significant yield advantage. Soil microbes play major roles in soil nutrient cycling and were affected by intercropping plants. This experiment was carried out to evaluate the changing of rhizosphere microbial community composition, and the relationship between microbial community and soil enzymatic activities, soil nutrients in maize/peanut intercropping system under the following three treatments： maize （Zea mays L.） and peanut （Arachis hypogaea L.） were intercropped without any separation （NS）, by half separation （HS） using a nylon net （50 μm） and complete separation （CS） by using a plastic sheet, respectively. The soil microbial communities were assessed by phospholipid fatty acid （PLFA）. We found that soil available nutrients （available nitrogen （Avail N） and available phosphorus （Avail P）） and enzymatic activities （soil urase and phosphomonoesterase） in both crops were improved in NS and HS treatments as compared to CS. Both bacterial and fungal biomasses in both crops were increased in NS followed by HS. Furthermore, Gram-positive bacteria （G＋） in maize soils were significant higher in NS and HS than CS, while the Gram-negative （G-） was significant higher in peanut soil. The ratio of normal saturated to monounsaturated PLFAs was significantly higher in rhizosphere of peanut under CS treatment than in any other treatments, which is an indicator of nutrient stress. Redundancy analysis and cluster analysis of PLFA showed rhizospheric microbial community of NS and HS of both plants tended to be consistent. The urase and Avail N were higher in NS and HS of both plants and positively correlated with bacteria, fungi （F） and total PLFAs, while negatively correlated with G＋/G- and NS/MS. The findings suggest that belowground interactions in maize/peanut intercropping system play important roles in changing the soil microbial composition and the dominant microbial species, which was closely related with the improving of soil available nutrients （N and P） and enzymatic activities.

Effect of Application of a Bacteria Inoculant and Wheat Bran on Fermentation Quality of Peanut Vine Ensiled Alone or with Corn Stover

To find an effective method for ensiling peanut vine （PV）, fermentation characteristics and nutritional values of PV silage and the mixture of PV with corn stover （CS） silage in a ratio of 1 ： 1 fresh weight, prepared by adding lactic acid bacteria （LAB）, 10% wheat bran （WB） and LAB＋WB at ensiling were evaluated in 2009 and 2010. The fermentation qualities of PV silage ensiled with the LAB and WB additives were improved compared with those of the control （PV ensiled alone）. However, the pH did not decline to the critical level of 4.2, and the nutritional values of the silage were not protected against losses in the LAB and WB addition silages. Ensiling PV in mixture with CS generated optimal moisture content and buffering capacity （BC） of ensiled materials. After adding the LAB and WB additives to mixture silage, especially adding LAB＋WB, the fermentation qualities and nutritional values of the mixture silage were improved significantly （P〈0.05）, and the Flieg＇s score reached to 99. The result suggested that it is a feasible method to ensile the mixed materials of PV with CS by adding LAB and high concentration of water soluble-carbohydrate materials for providing a good fermentation quality of PV silage.

QTL mapping of quality traits in peanut using whole-genome resequencing

Oil and protein content and fatty acid composition are quality traits in peanut.Elucidating the genetic mechanisms underlying these traits may help researchers to obtain improved cultivars by molecular breeding.Whole-genome resequencing of a recombinant inbred population of 318 lines was performed to construct a high-density linkage map and identify QTL for peanut quality.The map,containing 4561 bin markers,covered 2032 c M with a mean marker density of 0.45 c M.A total of 110 QTL for oil and protein content,and fatty acid composition were mapped on the 18 peanut chromosomes.The QTL q A05.1 was detected in four environments and showed a major phenotypic effect on the contents of oil,protein,and six fatty acids.The genomic region spanned by q A05.1,corresponding to a physical interval of approximately 1.5 Mb,contains two SNPs polymorphic between the parents that could cause missense mutations.The two SNP sites were employed as KASP markers and validated using lines with extremely high and low oil contents.These sites may be useful in the marker-assisted breeding of peanut cultivars with high oil contents.

不同气象条件对高油酸花生产量的影响

以河南、山东等5个花生主要产区培育的19个高油酸花生新品种为材料,于2019年~2021年连续3年在郑州种植,采集3年间花生关键生育期(6月~9月)的旬平均气温、旬降水量和旬日照时数等气象数据,结合各个品种的产量数据进行综合分析。研究结果表明,不同的品种对于气温、降水量和日照时数的敏感程度不同,冀花16、冀花11对开花下针期的温度和日照时数比较敏感,该时期的异常高温会致使其产量明显下降。中花413、中花412和花育917的产量与旬降水量成显著负相关,耐涝程度较差。对品种的稳定性分析表明,开农1768、南充1704、冀农16、开选016、花育961、郑农花25、郑农花23、冀农11、花育633和1508G等10个高油酸品种为年际间稳定性较好的品种,研究结论为高油酸品种的培育及深入利用提供了指导。

Effects of UV irradiation treatment on quality of peanut oil in storage life

Ultraviolet （UV） irradiation is proven to be an effective method to reduce aflatoxin in peanut oil, but the changes of peanut oil quality are not clear, especially in storage life. In this study, impacts of UV irradiation treatment on quality of peanut oil were investigated. Acid value, iodine value, oxidative stability and total phenol content that refer to rancidity and oxidation of oil were estimated under 365 nm UV irradiation processing and during different storage time. Results indicated that acid value was increased slightly under irradi-ation and in storage life while iodine value, total phenol content and oxidative stability de-clined weakly. Changes of these quality indexes were all within safe and acceptable levels. It can be confirmed by this study that UV irradiation is a safe strategy for AFB1 detoxification in peanut oil and not influence upon quality and stability of peanut oil.

Processing impact on volatile compounds profile in higholeic and normal peanut oil

With the improvement of living quality, high-oleic peanuts have drawn people’s attention. Increasing oleic acid content of peanut oil should be desired for improving shelf life and potential health benefits. The flavor is among the most important quality attributes for commercial acceptance of peanuts products. The volatile components of high-oleic pea-nuts oil and normal-oleic lines were compared in this research. First, volatile components of peanut oils were analyzed and identified by solid-phase microextraction/gas chroma- tography/mass spectrometry （SPME/GC/MS）. Then principal components analysis （PCA） was used to identify principal volatile components in peanut oils of different cultivars fo卜 lowing three different processing methods, including roasting, boiling and roasting, and roasting by microwave-assisted. The results showed that there were obvious differences of the volatile components between high-oleic and normal peanut oils. Among different pro-cessing method of peanuts, there was no clear separation on volatile components observed between roasting and boiling and roasting by microwave-assisted. Moreover, the same genotype had the similar flavor characteristic. Different processing methods had no signifi-cant effect on volatile components in peanut oils.

近红外光谱法快速检测花生籽仁主要品质指标定量模型的研究

建立花生籽仁粗蛋白、粗脂肪、油酸、亚油酸含量的近红外光谱快速测定方法。利用波通DA7200型近红外分析仪采集近红外光谱,采用凯氏定氮法、索氏提取法分别测定粗蛋白、粗脂肪的含量,采用气相色谱法测定油酸、亚油酸的相对含量,采用偏最小二乘法,构建花生籽仁主要品质指标含量的近红外预测模型。结果表明,模型对花生籽仁粗蛋白、粗脂肪、油酸、亚油酸含量的决定系数分为0.9270、0.9647、0.9915、0.9915,均方根误差分别为0.8702、0.5631、1.6671、1.4040。经外部验证,独立测试集决定系数分别为0.9608、0.9460、0.9605、0.9492。该模型对花生籽仁粗蛋白、粗脂肪、油酸、亚油酸含量的预测准确,可实现花生籽仁主要品质指标的快速、无损测定,提升高品质花生新品种的育种效率。

花生壳微晶纤维素改性聚乳酸的性能及机理

花生壳采用碱解、酸解、脱色等处理后制得微晶纤维素(MCC),将其作为填料加入到聚乳酸(PLA)中,通过溶液流涎成膜的方法制得不同MCC含量的MCC/PLA复合膜。分析了碱解过程中氢氧化钠浓度对MCC收率的影响、MCC含量对MCC/PLA复合膜性能的影响及其作用机理。利用傅里叶变换红外光谱仪(FTIR)、差示扫描量热仪(DSC)、扫描电子显微镜(SEM)、万能力学试验机等对MCC及MCC/PLA复合膜结构及性能进行表征和分析。结果表明,花生壳通过碱解、酸解、脱色等处理,将木质素等物质大量去除,制得MCC;MCC的加入,影响了PLA结晶结构的规整度及完善程度,降低了复合膜的熔融温度;随着MCC含量的增加,MCC/PLA复合膜的拉伸强度和断裂伸长率呈先增加后降低的趋势,均大于纯PLA的拉伸强度和断裂伸长率。当MCC的含量为5.6%时,MCC/PLA复合膜的拉伸强度为36.5 MPa,断裂伸长率达到最大值1.7%,与纯PLA相比,分别提高了52.1%和30.8%;SEM结果表明,该含量的MCC在PLA中分散均匀,复合膜光滑平整。

高油酸食用型黑花生新品种浙黑甜1号的选育

浙黑甜1号是浙江省农业科学院作物与核技术利用研究所以花育51号为母本、云南四粒黑为父本进行有性杂交,经过多年多代选育出的高油酸食用型黑花生新品种。该品种具有高产稳产、早熟性好、抗病性强、黑色种皮、口感微甜等突出特性。2020—2021年连续2 a参加浙江省区域试验,荚果平均产量为3 892.95 kg·hm^(-2),籽仁平均产量为2 817.30 kg·hm^(-2),分别比对照小京生增产30.37%和36.27%。浙黑甜1号蛋白质含量27.1%,粗脂肪含量45.8%,油酸含量76.9%,油亚比(O/L)12.05,抗叶斑病、青枯病,中抗锈病。该品种于2023年12月通过国家非主要农作物品种登记,适宜在长江中下游生态区浙江春播及夏播种植。