Detection of seven phytohormones in peanut tissues by ultra-highperformance liquid chromatography-triple quadrupole tandem mass spectrometry

Development of highly sensitive and reliable method for detection of phytohormones is of great significance to study plant hormones and agricultural production.In this study,an ultra-high-performance liquid chromatography-mass spectrometry/mass spectrometry method was established for separation and quantification of trans-zeatin,trans-zeatin riboside,gibberellin A_3,indol-3-acetic acid,salicylic acid,abscisic acid,and jasmonic acid(JA) without any label.The sepa ration was performed on an Agilent Explus Plus C18 column by using methanol and water as mobile phases with gradient elution.The target compounds were confirmed and quantified by mass spectrum via positive electrospray ionization for trans-zeatin,transzeatin riboside,indole-3-acetic acid,and via negative electrospray ionization for gibberellin3,salicylic acid,abscisic acid,and JA.The limits of detection ranged from 0.0127 ng L~(-1) for gibberellin A_3(GA_3) to 33.26 ng L~(-1) for JA and were lower than the currently reported values in literature.The proposed method was applied for qualitative and quantitative analyses of phytohormones in peanut gynophores and pods.The recoveries of the spiked phytohormones ranged from 80.20 to102.56%.The contents of seven endogenous hormones varied specifically in different development stages of peanuts.This study provides a highly sensitive and selective detection method for hormones and elucidates the growth and development of the gynophore and peanut fruit,which are controlled by seven endogenous hormones.     

海南甘薯主栽品种再生体系的优化

本研究以海南甘薯主栽品种‘心香’、‘广薯87’、‘川山紫’、‘宁紫薯1号’、‘薯绿一号’、‘高系14’和‘三角宁’为材料,从外植体的选择、不同消毒方案、培养基配方和防褐化剂筛选等方面进行研究,建立一套适合热带地区甘薯主栽品种的脱毒快繁优化再生体系。结果表明：侧芽增殖率与存活率最高;在不同的外植体消毒处理中,依次用75%酒精消毒60 s,2% NaClO消毒15 min,以及0.1% HgCl₂消毒15 min的污染率最低;最佳的生根培养基配方为：MS+0.05 mg/L NAA+0.1 mg/L GA₃;茎长生长最快的培养基配方为：MS+0.1 mg/L NAA+1 mg/L 6-BA+0.5 mg/L GA₃;干重增长最快的培养基配方为：MS+0.1 mg/L NAA+1.5 mg/L 6-BA+0.5 mg/L GA₃;在培养基中加入5~6 g/L硫代硫酸钠和1.25 g/L聚乙烯吡咯烷酮能有效地抑制外植体的褐变。     

大球盖菇1号的分离纯化及其液体培养基优化

为缩短大球盖菇(Stropharia rugoso-annulata)1号液体种生长周期,提高菌丝体产量,为大球盖菇液体种工业化生产提供参考和依据。试验以大球盖菇1号菌丝球干质量(Y)为指标,通过Plackett-Burman设计试验,得出玉米粉、MgSO4和VB1对大球盖菇的生物量有显著影响,通过最陡爬坡试验和Box-Behnken响应面分析法对大球盖菇液体培养基组成进行优化,得到大球盖菇1号最适液体培养基为蔗糖20 g·L^-1、蛋白胨6 g·L^-1、酵母膏2 g·L^-1、KH2PO43 g·L^-1、MgSO44.56 g·L^-1、玉米粉4.25 g·L^-1、VB12.43 g·L^-1,通过对模型进行验证试验,大球盖菇1号菌丝生物量为1.2154 g·150-1mL-1,较基础培养基相比生物量提高36.2%,且数学模型的预测值与试验观察值相符,相对误差约为1.9%,证明响应面法优化大球盖菇1号液体培养基可行。     

玉米粉中伏马毒素FB1基体标准物质的评价

为填补我国在伏马毒素基体标准物质的空白,本试验研制了以玉米粉为基体的伏马毒素FB1标准物质,玉米样品经过筛、加标、冷冻干燥、磨粉、混匀、密封分装后,用超高效液相色谱-串联质谱(UPLC-MS/MS)法检验样品的均匀性、稳定性,幵联合多家实验室对玉米粉中伏马毒素FB1含量定值,同时分析样品的不确定度。样品均匀性经斱差分析法表明F值为1.42,小于临界值F_(0.05),且伏马毒素FB1含量在觃定时间6个月内无明显变化。结果表明,均匀性与稳定性均符合标准物质的要求。样品定值为1475.56μg kg~(–1),不确定度为169.98μg kg~(–1)。该标准物质可替代进口标准物质,用于伏马毒素检测过程中的仪器校准、实验室质量控制和操作人员的水平考核等。     

菜心游离小孢子培养高频胚诱导培养基优化

【目的】对菜心小孢子培养的诱导培养基进行优化,以提高胚胎诱导率,创制优良DH系应用于育种实践。【方法】设置不同蔗糖浓度和激素(6-BA和NAA)浓度组合的NLN诱导培养基,以10份不同基因型的菜心为试材,进行小孢子培养及出胚率分析,筛选最佳诱导培养基。【结果】7份材料诱导出胚状体,不同基因型间的胚胎诱导率差异显著,说明基因型是影响小孢子形成的主要因素;在无激素处理中,100 g/L蔗糖浓度NLN(NLN-10)小孢子胚诱导率最高,出胚材料诱导率在3.45～30.46胚/皿;NLN含量减半的1/2 NLN-10培养基处理出胚率约为NLN-10处理的2/3;130 g/L蔗糖浓度NLN处理出胚率显著降低,最高仅为14胚/皿,蔗糖浓度为160 g/L的NLN(NLN-16)处理未出现胚状体;在NLN-10中分别添加0.1 mg/L 6-BA和NAA,促进出胚效果最佳,出胚率最高的 Rt19006达46.93胚/皿,是不添加激素处理的1.5倍;当NLN-10 中6-BA和NAA浓度均达到0.2 mg/L时,胚胎生长发育受到抑制。【结论】筛选出可有效促进菜心小孢子出胚的诱导培养基配方,即100 g/L蔗糖浓度的NLN+6-BA 0.10 mg/L +NAA 0.10 mg/L。     

一株盐芽孢杆菌的筛选及其粗酶的酶学特性研究

从浙江舟山市双峰盐场中筛选出一株分泌蛋白酶的嗜盐菌,结合16S rDNA基因序列鉴定、菌株形貌特征确定其为特氏盐芽孢杆菌(Halobacillustrueperi)命名为HalobacillustrueperiB1。经研究,该菌株最适生长NaCl浓度为5%,表明其为中度嗜盐菌。该蛋白酶在50℃和pH 7.5条件下具有最高的蛋白酶活力。对发酵培养时间的优化实验表明发酵8 h后B1分泌的蛋白酶活力最大。对发酵培养基的优化结果表明,脱脂奶粉为最适氮源和碳源;75 g·L~(-1)为最佳发酵盐浓度,对钙镁离子的依赖不明显。确定优化后的发酵培养基:Na HCO_30.06 g,KCl_2.0g,CaCl_2·2H_2O0.48 g,Mg SO_4·7H_2O 1.0 g,FeCl_3 0.001 g,NaCl 75.0 g,脱脂奶粉10.0 g。在初始pH 7.5,温度为40℃条件下发酵8 h。     

樟子松嫩枝扦插技术

研究了樟子松嫩枝扦插生根的特点以及插穗生根与母树年龄、扦插基质、扦插时间、生根促进剂质量分数、温度的关系。结果表明:①樟子松属于愈伤组织生根型,只有极少数为皮部生根,生根时间为2～4个月。扦插期间地表温度过低(低于20℃)不利于生根。②扦插的最好时间为6月上中旬,此时扦插的穗条处于半木质化时期。③樟子松扦插生根的最佳组合为:蛭石基质,插穗为2年生母树的穗条,生根促进剂为L2号。母株年龄对樟子松扦插生根有极为显著的影响,母树的年龄效应是樟子松穗条扦插生根的主要限制因子,2年生到5年生的插穗生根率从90%降到2.5%;而扦插基质之间和生根促进剂之间对生根率没有显著影响。基质对插穗生根影响的顺序从大到小为蛭石,草炭,珍珠岩 草炭(1∶1),河沙。生根促进剂以质量分数为1×10-4L2号最好,生根率达到67%,质量分数太小(0.5×10-4)或太大(3×10-4、5×10-4)都使得成活率下降。3种处理对生根率影响的顺序从大到小顺序为:插穗的母树树龄,扦插基质,生根促进剂。     

Theonellamides J and K and 5-cis-Apoa-theopalauamide,Bicyclic Glycopeptides of the Red Sea Sponge Theonella swinhoei

Theonella swinhoei is a fairly common inhabitant of reefs throughout the Indian and Pacific Oceans. Metabolomic analyses of samples of T. swinhoei collected in different depths in the Gulf of Aqaba revealed two chemotypes differing in the profiles of the theonellamides they produce, some of which seem to be unknown. Driven by this finding, we examined a sample of T. swinhoei collected more than 40 years ago in the southern part of the Gulf of Aqaba. Large-scale extract of this sample yielded four theonellamides, the known theopalauamide (4), as the major component, and three new metabolites, theonellamide J (1), 5-cis-Apoa-theopalauamide (2), and theonellamide K (3), as the minor components. The planar structure of these complex cyclic glycopeptides was elucidated by combination of 1D and 2D NMR techniques and HRESIMS. The absolute configuration of the amino acids was established by Marfey’s and advanced Marfey’s methods, and the absolute configuration of its galactose unit using “Tanaka’s method” for monosaccharides. The biological activity of the pure compounds was tested for antibacterial activity and for cytotoxicity to HTC-116 cell line. The compounds presented significant cytotoxicity against the HTC-116 cell line, illuminating the importance of the Apoa subunit for the activity.     

5种动物源食品中的氟虫腈及其3个代谢物残留量检测

利用QuEChERS技术结合超高效液相色谱-串联质谱的方法检测5种动物源食品(鸡肉、鸡蛋、猪肝、牛奶、猪肉)中氟虫腈及其3个代谢物残留量。样品用乙腈提取,经低温处理,多壁碳纳米管(MWCNTs)、弗罗里硅土(Florisil)和C18分散固相萃取(d-SPE)净化,外标法定量。在不同浓度的加标水平下,氟虫腈及其代谢物在5种动物源食品中的平均回收率为72.1%～113.8%,相对标准偏差RSD为1.9%～17.6%,检出限LOD在0.73～1.94μg/kg范围内,定量限LOQ在5～10μg/kg范围内。该方法具有简单、灵敏、准确等优点,适用于动物源食品中氟虫腈及其代谢物的快速筛查和定量检测。     

敌百虫对三疣梭子蟹幼体的急性毒性研究

试验结果表明,随着三疣梭子蟹幼体的发育,其半致死质量浓度以及安全质量浓度呈上升趋势。三疣梭子蟹Ⅰ期溞状幼体24 h LC50和48 h LC50分别为0.0095 mg/L和0.0082 mg/L,安全质量浓度为0.0018 mg/L;三疣梭子蟹Ⅱ期溞状幼体24 h LC50和48 h LC50分别为0.019 mg/L和0.010 mg/L,安全质量浓度为0.0038 mg/L;三疣梭子蟹Ⅳ期溞状幼体24 h LC50和48 h LC50分别为0.085 mg/L和0.047 mg/L,安全质量浓度为0.0043 mg/L;三疣梭子蟹大眼幼体24 h LC50和48 h LC50分别为0.085mg/L和0.050 mg/L,安全质量浓度为0.0052 mg/L。