Research of Seed Testa Structure and Storage Material of Peanut Germplasm with Different Resistance to A. flavus

There was an obvious relationship between seed testa structure, storage material and resistance to A. flavus of peanut. Results showed that seed testa of peanut germplasm with high resistance （HR） to A. flavus infection had thicker wax layer, integrated and tight epidermis layer, regular vascular tissue range. However, the seed testa of peanut germplasm with high sensitivity （HS） to A. flavus had the reverse results, and results of those with medium resistance （MR） to A. flavus lay in between, but changes of testa thickness were not significant among different resistance kinds. Results also showed that some seed storage materials were closely related with resistance potential to A. flavus. It seemed that varieties with higher resistance to A. flavus had higher oleic acid and protein content, lower linoleic acid and fat content. Content of palm acid, total sugar and VE did not show positive relationship with the resistance to A. flavus.

Peanut testa extracts enhance anticancer effect of cisplatin against human cholangiocarcinoma cells via modulation of histone deacetylase inhibitory activity

Objective:To investigate the effect of combination treatments of cisplatin and KK4 and ICG15042 peanut testa extracts against cholangiocarcinoma cells in vitro.Methods:The growth inhibition,cell cycle arrest and apoptosis of cholangiocarcinoma cells were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and flow cytometry analysis,respectively.The levels of proteins involved in apoptosis were assessed using Western blotting assays.The caspase activity was assessed using a colorimetric caspase activity assay.Results:Cisplatin and peanut(KK4 and ICG15042)testa extracts inhibited the growth of cholangiocarcinoma cell lines(KKUM214 and KKU-100 cells)in a dose-and time-dependent manner.The combination treatments reduced cell viability and induced apoptosis of cholangiocarcinoma cells more efficiently than singledrug treatments.Cancer cell death synergistically mediated by cisplatin and peanut testa extracts was observed in KKU-M214 cells(combination index<1.0)but not in KKU-100 cells(combination index>1.0).The combination treatments also increased the subG1 population and caused KKU-M214 cell cycle arrest at S and G2/M phases,which were the combined effects of cisplatin(S phase arrest)and peanut testa extracts(G2/M phase arrest).In addition,p ERK1/2,Ac-H3,Bcl-2 and proteins related to apoptosis,including Bax and caspases 3,8,9,exhibited enhanced expression in KKUM214 cells.The combination treatments caused down-regulation of p53,whereas the expression of p21 was fairly constant when compared with cisplatin single drug treatment.Conclusions:Peanut testa extracts in combination with cisplatin synergistically reduce cell viability and induce apoptosis through stimulation of caspases 3,8 and 9 in KKU-M214 cells.

Phytochemicals Screening, Phenolic Estimation and Evaluation for Anti-Oxidant, Anti-Inflammatory and Anti-Microbial Activities of Sequentially Soxhlet Extracted Coconut Testa

Background: In many coconut industries, the outer layer of thin brown skin of coconut kernel known as testa is peeled out as a byproduct. Despite the testa is rich in fat and plenty of polyphenolic compounds, it has been underutilized either as animal feed, serving as raw materials for bio-diesel production or discarded directly. Anticipating coconut testa (CT) as a natural source of multiple phyto-chemicals, its exploitation for the pharmacological activity or utilization as value added product is required which may reduce the disposal costs as well. Methods: Secondary metabolites from CT were extracted sequentially with different organic solvents based on polarity in the soxhlet apparatus followed by extraction with sterilized water. The crude dried extracts thus prepared were evaluated for qualitative screening of phytochemicals and quantitative estimation of total phenols, flavonoids and tannin content. Moreover, the antioxidant, anti-inflammatory and anti-microbial activities were also investigated. Results: Phytochemicals screening revealed the presence of polyphenolic compounds in methanolic fraction including phenols (822.60 ± 16.36 mg/g), flavonoids (103.30 ± 9.78 mg/g) and tannin (663.50 ± 19.26 mg/g), whereas non-phenolic compounds were present in other fractions. While methanolic fraction showed invariably the highest anti-oxidant activity in multiple assay methods, non-phenolic compounds in aqueous and chloroform fractions exhibited high anti-inflammatory activity. Antimicrobial activity was observed by both phenolic and non-phenolic compounds. Conclusion: The findings of the study reveal that CT is a rich source of various polyphenolic and non-phenolic natural antioxidants, anti-inflammatory and antimicrobial compounds. These findings are promising and form the basis to identify the number of active components and their characterization.

Phenolic compounds from Peanut testa

Phytochemical investigation of peanut testa(the seed coat of Arachis hypogaea L.)led to the isolation of eight phenolic compounds,including caffeic acid(1),methyl caffeate(2),ethyl caffeate(3),methyl protocatechuate(4),ethyl protocatechuate(5),butyl protocatechuate(6),(E)-p-hydroxycinnamic acid methyl ester(7),and resveratrol(8).The structures of the compounds were elucidated through spectroscopic data analysis and comparison with the previously reported literature.Among them,compounds 2,3,5,and 6 were obtained from Arachis hypogaea L.for the first time.

Effects of Transparent Testa8(TT8) gene and Homeobox12(HB12) gene silencing in alfalfa(Medicago sativa L.) on molecular structure spectral profile in relation to energy,degradation,and fermentation characteristics in ruminant systems

Alfalfa(Medicago sativa L.) is a legume forage that is widely cultivated owing to its high biomass yield and favorable nutrient values. However, alfalfa contains relatively high lignin, which limits its utilization.Downregulation of two transcriptional factors, Transparent Testa8(TT8) and Homeobox12(HB12), has been proposed to reduce lignin content in alfalfa. Therefore, silencing of TT8(TT8i) and HB12(HB12i) in alfalfa was achieved by RNAi technology. The objective of this project was to determine effect of gene modification through silencing of TT8 and HB12 genes in alfalfa plants on lignin and phenolic content,bioenergic value, nutrient supply from rumen degradable and undegradable fractions, and in vitro ammonia production in response to the silencing of TT8 and HB12 genes in alfalfa. All gene silenced alfalfa plants(5 TT8i and 11 HB12i) were grown under greenhouse conditions with wild type as a control.Samples were analyzed for bioactive compounds, degradation fractions, truly digestible nutrients, energetic values and in vitro ammonia productions in ruminant systems. Furthermore, relationships between physiochemical, metabolic and fermentation characteristics and molecular spectral parameters were determined using vibrational molecular spectroscopy. Results showed that the HB12i had higher lignin, while TT8i had higher phenolics. Both silenced genotypes had higher rumen slowly degraded carbohydrate fractions and truly digestible neutral detergent fiber, but lower rumen degradable protein fractions. Moreover, the HB12i had lower truly digestible crude protein, energetic values and ammonia production compared with other silenced genotypes. In addition, in relation to the nutritive values of alfalfa, structural carbohydrate parameters were negatively correlated, whereas alpha/beta ratio in protein structure was positively correlated. Furthermore, good predictions were obtained for degradation of protein and carbohydrate fractions and energy values from molecular spectral parameters. In conclusion, silencing of the TT8 and HB12 genes decreased protein availability and increased fiber availability. Silencing of the HB12 gene also increased lignin and decreased energy and rumen ammonia production. Moreover, nutritional alterations were closely correlated with molecular spectral parameters. Therefore, gene modification through silencing the TT8 and HB12 genes in alfalfa influenced physiochemical, metabolic and fermentation characteristics.

白果外种皮中银杏酚酸的中试生产调试及其稳定性试验研究

目的:探索白果外种皮中银杏酚酸的中试生产调试及其稳定性试验研究。方法:在确定白果外种皮中银杏酚酸的最佳提取参数、有机溶剂萃取纯化参数以及大孔树脂柱层析纯化参数后完成中试调试,并进行银杏酚酸的稳定性研究。结果:在确定的白果外种皮中银杏酚酸提取纯化工艺下,平均每批烘干白果外种皮投料量为20 kg,可以获得银杏酚酸2.43 kg,产率为11.98%,银杏酚酸含量为68.38%,回收率为75.76%。在参照2020版《中国药典》对提纯产品进行了银杏酚酸的稳定性试验,确定银杏酚酸的包装方式为密封避光保存,首先采用圆底封盖塑料管密封包装,再用自封袋进行二次包装,最后在最外层采用塑封袋进行包装。由于长期试验时限不足,目前初步得出该包装下银杏酚酸在6个月内含量稳定。结论:该研究将白果外种皮中银杏酚酸的回收利用生产工艺进行中试调试并对银杏酚酸的稳定性进行研究,为银杏产业中银杏酚酸的产业化实施提供参考数据。

去红衣与超声处理对花生油体提取及其乳化特性、抗氧化活性的影响

为了对花生油体的工业化提取和基于油体乳化体系新产品的开发提供参考,探究了去红衣与超声处理对花生油体提取及其乳化特性、抗氧化活性的影响。不同预处理的花生经水相提取,得到油体、清液和沉淀三相,测定了三相中固形物、脂肪、蛋白质的含量和分布,并分析了油体的乳化特性、总酚与总黄酮含量以及抗氧化活性。结果表明:去红衣将脂肪在油体中的分布由63.36%提高至65.30%,与超声联合处理时进一步将其提高至68.13%;去红衣处理不利于花生油体的稳定性和抗氧化活性;与对照组相比,去红衣组油体乳液平均粒径由2510.00 nm上升至2953.67 nm,动力学稳定性降低;去红衣组油体总酚、总黄酮含量最低,分别为71.01μg/g与47.14μg/g;超声处理促进了花生油体的溶出,且提取的油体乳化稳定性和抗氧化活性最高,其乳液平均粒径为1742.00 nm,乳化活性指数和乳化稳定性指数分别为114.48 m^(2)/g与1848.40 min,动力学稳定性最佳,总酚、总黄酮含量分别为101.24μg/g与59.97μg/g。综上,去红衣处理有利于花生油体的提取,但会降低其稳定性与抗氧化活性,而超声处理不仅有利于花生油体的提取,还增强了其乳化稳定性和抗氧化活性。

两种大豆籽粒萌发特性差异的比较转录组分析

大豆种子的萌发决定着幼苗建成和生长发育进程,快速萌发及高发芽率是大豆生产中获得高产的重要保障。本研究以不同萌发特性的黑农35及其诱变株系M为材料,基于萌发初期种皮微形态观察和主要生理指标检测,拟通过“露白期”(吸水后24 h)转录组的差异基因分析,进一步明晰大豆种子萌发机理。结果表明,两者的吸水率和发芽率存在显著差异;扫描电镜观察发现,与黑农35相比,M的种脐较长,种皮较薄,这些形态特征可能是萌发初期M吸水更快的主要原因。萌发过程中脱落酸(ABA)含量逐渐下降,可溶性蛋白含量均呈先下降后上升趋势,可溶性糖含量均为上升-下降-上升的“S”形。转录组分析发现两者共有1122个差异基因,其中478个上调,644个下调,差异基因主要富集在光合作用途径,筛选到22个与光合途径相关的候选基因,在M中均呈下调趋势,可能引起光形态建成较慢,进而导致诱变株系M萌发率变低,研究结果可为解析大豆种子萌发的调控机制提供思路。

芍药属组间杂交种皮开裂种子的解剖观察与胚培养

【目的】探究芍药属组间杂交种皮开裂种子的内部状态和胚培养成苗表现,为揭示种皮开裂种子的成苗潜力、提高组间远缘杂交育种效率提供参考。【方法】以24个芍药属组间杂交组合种皮开裂的种子为试验材料,解剖后观察并统计胚和胚乳的发育情况,通过离体培养观察胚的萌发和生长规律,分析不同组合种皮开裂种子发育和成苗差异。【结果】芍药属24个组间杂交组合中均出现了种皮开裂的杂交种子,平均种皮开裂率高达61.08%;1767粒种皮开裂种子中,有11.09%的胚乳已腐烂;胚乳未腐烂种子中,约90%以上种子的胚乳存在不同程度的损伤或病变,24.18%的种子内部未观察到可见胚。对解剖得到的1185个组间杂种胚进行离体培养发现,有24.73%的胚不萌发,63.47%的胚生长异常,仅有4.47%的胚能正常生长,得到完整植株。胚培养得到的完整植株具备良好的活力,移栽成活率达到86.79%。24个杂交组合种皮开裂种子通过离体胚培养共得到46株移栽成活的组间杂种苗,平均成苗率为2.60%,接近正常组间杂交种子的播种成苗率(2.75%)。【结论】种皮开裂现象在芍药属组间杂交种子中普遍存在,是组间远缘杂交障碍的重要表现之一,用离体胚培养技术能有效克服这类障碍,提高成苗率。

In vitro anti-Helicobacter pylori activity of aqueous extract from Persian Oak testa

Objective:Growing problem of antibiotic resistance in Helicobacter pylori,as a common cause of chronic gastritis and even stomach cancer,demands searching for novel candidates of herbal sources.This study is aimed at assessing the antimicrobial activity of aqueous extract obtained from Quercus brantii var.persica seed coat(Testa)on H.pylori isolated from gastric biopsy specimens.Methods:Such specimens were collected from 100 patients presenting with endoscopic gastroduodenal findings.Testa extracts were prepared from Persian Oak forests in the province of Kohgiluyeh and BoyerAhmad,IRAN.H.pylori isolates were obtained by a series of standard bacteriology tests and cell culture,then were confirmed by PCR.The activity of testa extracts towards 25 H.pylori isolates was assessed by well diffusion method,microdilution assay,and a disk diffusion assay in vitro.Results were analyzed statistically by one-way ANOVA analysis.Results:Aqueous extract of testa demonstrated an antimicrobial activity with zone diameters of inhibition ranged from 0 mm to 40 mm.Its inhibitory activity increased simultaneously with increasing extract concentration.The lowest MIC and MBC were both recorded as 2μg/m L.Anti-H.pylori activity of testa extract was approximately close to tetracycline and metronidazole and less than amoxicillin.A potent extract of testa possessed significant inhibitory activity(P<0.05).Conclusion:Testa extract is suggested as a natural therapeutic source against the gastric H.pylori infection.However,evaluating the in vivo activity of this extract is necessary too.

盐胁迫下不同花生品种的类黄酮含量和抗氧化酶活性

花青素等黄酮类物质,既是人体所需的营养保健成分,也是介导植物适应逆境胁迫的重要代谢物。以3个不同种皮颜色的花生品种为研究对象,在苗期以150 mmol/L的NaCl进行盐胁迫处理,测定植株性状、类黄酮含量和抗氧化酶活性,分析不同品种的抗氧化能力和耐盐性。结果表明,盐胁迫抑制了株高、叶面积和生物量,3个品种的耐盐系数从高到低依次为济花黑1号、济花红1号、远杂9102。与远杂9102相比,济花红1号和济花黑1号MDA含量相对较低。盐胁迫显著提高了济花红1号和济花黑1号根系的SOD、POD、CAT等抗氧化酶活性。济花红1号和济花黑1号根系类黄酮含量和在盐胁迫下的相对值大于远杂9102。相关分析表明,相对类黄酮含量和MDA含量与耐盐系数显著相关,类黄酮与SOD活性、MDA含量、株高和叶面积相对值极显著相关。盐胁迫引起济花红1号和济花黑1号类黄酮大量积累,激活了抗氧化保护能力,可有效降低MDA的积累,减轻氧化损伤,缓解盐胁迫对植株生长的抑制作用。研究结果为筛选和推广耐盐碱的彩色花生提供了依据。

正丁醇-盐酸法测定花生红衣中原花青素的方法优化

以花生红衣为原料,以原花青素含量为指标,采用优化后的正丁醇-盐酸法测定花生红衣提取物中原花青素的含量。通过单因素试验确定液料比、烘箱温度、保温时间对原花青素含量的影响。采用响应面软件,以单因素试验为基础,设计三因素三水平的响应面试验,并对试验结果进行分析。结果表明,影响花生红衣中原花青素含量的因素顺序为液料比>烘箱温度>保温时间,根据实际试验可行性,优化后的正丁醇-盐酸法的最佳条件分别为液料比65∶1(mL/g)、烘箱温度90℃、保温时间60 min,在此条件下原花青素的含量达到最高为135.003 mg/g。该方法的加标回收率为97.48%~98.78%,相对标准偏差为0.4%~1.1%。优化后的正丁醇-盐酸法具有较高的准确度和精密度,可为原花青素含量的测定方法优化提供参考。

拟南芥种皮色素形成突变体的筛选与表型鉴定

类黄酮在植物应答各种环境胁迫和种皮发育调控中起着重要作用。通过甲基磺酸乙酯(EMS)诱变筛选获得1个透明种皮突变体,与野生型拟南芥(Arabidopsis thaliana)(Col-0)相比,突变体成熟的种子颜色为黄色,其表型性状由隐性单基因控制。利用图位克隆和精细定位技术将突变基因定位于5号染色体MAH20的BAC上,是TT4(At5G13930)基因的第1 299位碱基C突变为T,使得第324位氨基酸甘氨酸突变为谷氨酸。TT4(transparent testa 4)编码1个类黄酮合成的结构基因查尔酮合酶(CHS),突变后种皮透明,种子颜色为黄色,突变体命名为tt4-1。利用功能回补突变体恢复褐色种皮表型,进一步证明了TT4在调节种皮颜色发育过程的重要作用。启动子偶联GUS基因组织表达分析显示TT4基因在植株幼苗的根、茎、叶和花中均有表达,生理表型分析结果显示与野生型相比,突变体tt4-1种子萌发早,幼苗主根短、侧根和根毛较多,成苗叶片气孔开度大和失水率高等特性。该研究将为进一步阐述TT4基因功能奠定理论依据。

基于欧标低温平衡环境型房间量热计

为了满足欧盟新的季节能效检测标准要求,需要开发出基于欧标的低温平衡环境型房间量热计进行空调季节能效测试。本项目根据量热计法测试原理;非稳态控制原理;小负荷能力测试原理等量热计工作原理,设计了试验室本体及气流组织,并根据EN14825标准中不同的试验工况要求,核算出试验室内外侧所需最大的冷热负荷。最终采用直接蒸发压缩制冷循环作为低温冷媒流量系统,选择了R404A作为制冷剂以及合适的冷冻机组合,围绕空气调节系统、测控系统、数据处理系统、实时监控系统等关键系统,开发出了各项指标达到欧盟标准测试要求的低温平衡环境型量热计。结论:本项目成果的应用,解决了空调器检测领域的重大关键问题,对推动空调器行业突破欧盟的技术壁垒发挥了重要作用。

柱型灯PP灯罩粉化性能研究

本文利用热氧老化建立PP粉化加速模型,通过热和蓝光叠加老化得到蓝光加速系数,并研究了等离子处理工艺对PP粉化寿命的影响,利用氧化诱导值、热氧粉化寿命等测试,确定等离子处理工艺的加速系数。结果表明,本试验所用PP材料在150℃热氧粉化寿命为2952h,蓝光辐照为46.59W.m-2时PP灯罩的150℃粉化寿命为950h,则蓝光加速系数为3.11;等离子空气压缩泵功率200W处理的PP灯罩150℃粉化寿命最长为1540h,等离子加速系数最小1.92;最终结合热氧加速模型和各因子加速系数模拟推算出30W柱型灯在正常使用(温升100℃)的粉化寿命为27145h,远大于产品设计15000h的寿命要求。

高粱单宁在认识上的谬误

1谬误1:所有高粱都含单宁事实:单宁仅存在于有色种皮的高粱品种中(图1)。种皮是否存在受B1_B2_基因控制。当B1_B2_为显性时,种皮就会有颜色。种皮没有颜色的高粱不含单宁,但是在分析检测的过程,一些非单宁物质也会吸收光波,并被误认为是单宁。

山杏种皮黑色素提取工艺研究

研究了提取溶剂及其浓度、提取温度和有无氮气保护及提取时间对山杏种皮黑色素提取效果的影响，并对提取的山杏种皮黑色素与人工合成的多巴黑色素的理化性质进行了比较。结果表明，该黑色素适宜于用0．5mol／L NaOH在60℃下提取2次，每次12h，提取宜在氮气下进行，以防止黑色素的过度氧化。在该工艺条件下，经过酸水解和进一步有机溶剂洗涤及反复沉淀纯化以后，可从5g山杏种皮中得到黑色素237mg，得率为4．73％；山杏种皮黑色素中总多酚含量较高，约为262mg／g。山杏种皮黑色素与合成多巴黑色素的理化性质非常相似，不溶于水和常见的有机溶剂，在1mol／L KOH溶液中完全溶解，pH小于3时发生沉淀，可被KMnO4、K2Cr2O7、NaClO和H2O2氧化漂白，多酚定性反应呈阳性；在紫外一可见光区没有吸收峰，红外光谱中在3400cm^-1 附近和1650～1620cm。各有一个强的吸收带。

花生红衣中多酚类物质清除DPPH自由基能力和抑菌性能的研究

研究了花生红衣多酚类物质对DPPH自由基的清除能力及抑菌性能。将花生红衣多酚粗提和纯化后的物质与没食子酸标准品、单宁酸标准品及抗坏血酸的EC50值作比较,得到花生红衣多酚粗提物EC50(0.194mg/L)<纯化花生红衣多酚EC50(0.705mg/L)<没食子酸EC50(0.760mg/L)<单宁酸的EC50(5.409mg/L)<抗坏血酸的EC50(3.745mg/L)。纯化后花生红衣多酚对枯草芽孢杆菌、大肠杆菌、青霉、黑曲霉和毛霉均有抑菌性能,最低抑菌浓度为250mg/L。

花生深紫色种皮颜色基因的遗传分析及SSR标记

以种皮呈深紫色的花生品种珍珠黑和粉红色品种粤油13的杂交后代F1-F3群体为材料,通过遗传分析和SSR分子标记探讨花生种皮颜色基因的遗传连锁规律。结果表明,花生深紫色种皮颜色受一对不完全显性主效基因控制,该基因与SSR标记“PM93/630-600”连锁,连锁距离为5.4 cM。

高粱胚乳细胞与母体组织发育关系的研究

【目的】探明高粱颖果发育过程中胚乳细胞与母体组织发育关系及其胞内淀粉体的发育特性。【方法】通过挂牌和记号笔点颖相结合的方法对供试高粱品种KS-304花后发育天数进行精确标记,详细跟踪观测了颖果的生长;采取Spurr树脂包埋、番红-甲基紫复染法制作半薄切片,光镜下详细观察了高粱颖果发育各过程中胚乳各部位以及种皮果皮结构的变化差异与联系;采用扫描电镜研究发育中后期及成熟时颖果断面不同部位细胞内淀粉体的形态;应用冰冻切片,荧光显微镜观察了成熟颖果横断面的结构。【结果】颖果发育分形成期、乳熟期、蜡熟期与完熟期,内胚乳发育分为游离核期、细胞化期、分化期、发育期及成熟期;其中颖果发育形成期与胚乳发育的前3个时期相对应,乳熟期对应发育期,而蜡熟期与完熟期对应胚乳的成熟期。颖果发育早期,珠心组织存留时间较长,珠心表皮细胞约在花后15 d消失。花后7 d,表层胚乳细胞开始积累脂质体,11 d时转为糊粉层细胞,成熟时糊粉层为1层,其细胞内除常规糊粉粒圆球体外,还含有少量单粒淀粉体,粒径约3μm。亚糊粉层细胞位于糊粉层细胞与内胚乳细胞之间,兼糊粉层与内胚乳细胞特点,贮藏大量蛋白体,细胞内淀粉体构成复杂。内胚乳发育存在区域差异,中央近胚处的细胞物质积累滞后于周缘胚乳细胞,成熟时,前者淀粉体充实较为疏松,发育为粉质胚乳,后者充实紧密,淀粉体彼此受到挤压呈多面体,最终发育为角质胚乳。高粱胚乳细胞内存在于不同于其他谷物颖果胚乳淀粉体的"发生中心"结构,主要表现为淀粉粒在管状质体内生长,随着体积的增长,后期与"发生中心"分离而形成新的淀粉体。果皮发育前期增厚,中后期呈缓慢减薄的趋势,中果皮发育后期细胞内观察到淀粉体存在特殊的二次增长,构成从前期复粒淀粉体为主转为单粒;"种皮"来源于珠心、珠被细胞依次降解后的胞壁残留堆叠而成,中间含有透明角质层。【结论】白高粱KS-304胚乳发育与玉米颖果胚乳类似,最终形成角质与粉质胚乳;颖果胚乳淀粉体发育存在独特的"发生中心";中果皮发育后期其细胞可能行使积累同化产物"库"的作用。