利用CRISPR/CAS9技术编辑水稻香味基因Badh2

【目的】香稻作为一类特殊的水稻群体,以其清香可口的品质特性备受消费者的欢迎。到目前为止,水稻中的香味主要受第8染色体上编码甜菜碱醛脱氢酶基因Badh2控制。【方法】通过CRISPR/CAS9技术对中花11的香味基因Badh2进行编辑。【结果】获得转基因T_0代植株并对其所衍生的T_1代20个单株进行了鉴定分析,获得了一个剔除了载体骨架且第1外显子上插入一个碱基T的突变体材料。该材料中Badh2 RNA水平显著下调;利用GC-MS技术测定野生型及突变体材料籽粒2-乙酰-1-吡咯啉含量,结果表明突变体材料中的香味物质显著增加;此外,我们还对野生型及T_2代香型植株水稻产量及稻米蒸煮食味品质进行了考查及测定分析,发现除分蘖数及结实率呈现出显著差异外(P0.05),其余各项指标在两组材料间都无显著差异。【结论】通过CRISPR/CAS9技术成功地对水稻香味基因进行了编辑,可为香稻育种提供丰富的理论指导,加快香稻的育种进程。     

土壤有机碳对沼渣或牛粪施用后温室气体排放潜力的影响—盆栽试验结果

A change in the European Union energy policy has markedly promoted the expansion of biogas production.Consequently,large amounts of nutrient-rich residues are being used as organic fertilizers.In this study,a pot experiment was conducted to simulate the high-risk situation of enhanced greenhouse gas (GHG) emissions following organic fertilizer application in energy maize cultivation.We hypothesized that cattle slurry application enhanced CO2 and N2O fluxes compared to biogas digestate because of the overall higher carbon (C) and nitrogen (N) input,and that higher levels of CO2 and N2O emissions could be expected by increasing soil organic C (SOC) and N contents.Biogas digestate and cattle slurry,at a rate of 150 kg NH4+-N ha-1,were incorporated into 3 soil types with low,medium,and high SOC contents (Cambisol,Mollic Gleysol,and Sapric Histosol,termed Clow,Cmedium,and Chigh,respectively).The GHG exchange (CO2,CH4,and N2O) was measured on 5 replicates over a period of 22 d using the closed chamber technique.The application of cattle slurry resulted in significantly higher CO2 and N2O fluxes compared to the application of biogas digestate.No differences were observed in CH4 exchange,which was close to zero for all treatments.Significantly higher CO2 emissions were observed in Chigh compared to the other two soil types,whereas the highest N2O emissions were observed in Cmedium.Thus,the results demonstrate the importance of soil type-adapted fertilization with respect to changing soil physical and environmental conditions.     

开垦年限对黑土氮初级转化速率和净转化速率的影响

以东北黑土区开垦2 a和开垦30 a的典型旱作土壤为研究对象,采用15N同位素成对标记技术开展室内培养试验,利用数值计算模型(FLUAZ)计算不同开垦年限土壤的氮初级转化速率,以比较不同开垦年限黑土氮初级转化速率和净转化速率的差异,明确开垦年限对黑土氮转化过程的影响。结果表明,与开垦2a土壤相比,开垦30a土壤的有机碳和水溶性有机碳含量显著降低,导致土壤氮初级矿化速率和初级固定速率也显著降低。但开垦30a土壤的初级硝化速率、净硝化速率和净氮矿化速率却显著高于开垦2a土壤。两个开垦年限土壤的初级硝化速率分别为净硝化速率的1.15倍和1.02倍,说明土壤微生物对硝态氮的固定很少。开垦30a土壤的m/i值(氮初级矿化速率与初级固定速率之比)和n/ia值(初级硝化速率与初级铵态氮固定速率之比)均显著大于1,而开垦2 a土壤的m/i值和n/ia值均接近1。表明开垦2 a土壤的氮矿化与固定过程紧密偶联,氮素损失的风险较小,而开垦30 a土壤中氮矿化量超过了固定量,这为硝化作用的进行提供了底物,增加了硝酸盐反硝化和淋溶风险。     

基于转录组测序的半滑舌鳎长链非编码RNA的初步鉴定与分析

以抗病家系与易感家系半滑舌鳎为材料,进行哈维弧菌感染实验,并对易感家系感染前(CsSU)、易感家系感染后(CsSC)、抗病家系感染前(CsRU)、抗病家系感染后(CsRC)4组进行转录组测序,根据RNA-seq数据挖掘半滑舌鳎长链非编码RNA信息;通过生物信息学分析,筛选出与抗哈维弧菌病相关的差异长链非编码RNA(long non-coding RNA, lncRNA)。结果显示,共识别出4 584个lncRNA座位,包含5 714个转录本;其基本特征与编码基因的比较分析,lncRNA的GC含量低于编码基因,单外显子基因数多于编码基因,转录本的平均长度长于编码基因,基因表达量低于编码基因。对4组样品进行两两比较(CsRU vs CsSU、CsRC vs CsSC、CsRC vs CsRU、CsSC vs CsSU)分别筛选出818、813、261、140个差异表达lncRNA,其中CsRU与CsSU之间、CsRC与CsSC之间lncRNA数目差异最多,通过聚类分析确定了各实验组的表达模式之间的联系,CsRU与CsSU之间的表达模式最为相近。通过共表达分析,预测出lncRNA和274个编码基因可能存在14 539种相互关系,并进行了功能注释,进而筛选出7个关键lncRNA。qRT-PCR结果显示,差异表达lncRNAs的表达模式和转录组数据得到的基本一致。研究结果为揭示lncRNA在半滑舌鳎抗哈维弧菌免疫调控反应中的作用提供重要的参考数据。     

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.     

不同浓度NAA/6BA配比对不同品种甘薯茎尖培养特性的影响

为明确不同品种甘薯茎尖培养的最佳NAA/6BA配比,设置0.1 mg/L/2.5 mg/L、0.2 mg/L/2.5 mg/L、0.2 mg/L/1.0 mg/L 3组NAA/6BA浓度配比处理,观测不同浓度激素处理对‘北京553’、‘红香蕉’、‘苏薯8号’、‘烟薯25’、‘安吉芋’、‘济徐23’、‘渝紫7号’、‘商薯19’茎尖培养成苗率、愈伤组织直径、不定根数目、叶片数和植株高度的影响,同时调查不同品种试管苗的移栽成活率。结果表明,‘红香蕉’、‘济徐23’、‘渝紫7号’、‘商薯19’在0.1 mg/L/2.5 mg/L的处理下培养效率最高;‘安吉芋’在0.2 mg/L/2.5 mg/L的处理下培养效率最高;‘苏薯8号’、‘北京553’、‘烟薯25’在0.2 mg/L/1.0 mg/L的处理下培养效率最高。因此,在甘薯茎尖培养过程中,不同甘薯品种适宜的NAA/6BA配比存在显著差异,在实际生产中应根据品种特性来进行调整激素的用量和比例。     

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。     

多酚类化合物协同抗氧化性研究进展

多酚类化合物是一种大量存在于植物体内的物质，因其具有优良的抗氧化、抗癌、抗衰老等 性能而受到研究者们的广泛关注。多项研究表明具有抗氧化性能的物质之间大多具有协同作用，两 种或两种以上的物质化合，其抗氧化能力往往强于单一物质的抗氧化能力。近年来，许多研究者将目 光聚焦到多酚类物质与小分子物质化合所具有的协同抗氧化能力的研究上并做出多项试验。本文论 述了多酚类化合物的抗氧化机理及协同抗氧化机理，并以茶多酚与维生素C、维生素E 化合为例对协 同抗氧化进行论述。     

浮性水产饲料膨化加工参数优化研究

为优化浮性水产饲料挤压膨化加工参数，以膨化度、容积密度、吸水性和溶失率作为饲料加工品质评价指标，采用响应面分析法研究了螺杆转速、出料段机筒温度和物料含水率对饲料加工品质的影响，并利用扫面电镜观测了饲料微观形貌。结果表明：适中的螺杆转速、出料段机筒温度以及低物料含水率，有利于形成较高的饲料膨化度；高螺杆转速与高出料段机筒温度有利于形成较低的饲料容积密度、吸水性以及溶失率；优化后的浮性水产饲料膨化加工参数为螺杆转速130 r·min^-1，出料段机筒温度150 ℃，物料含水率10%，其加工后的饲料膨化度、容积密度、吸水性和溶失率分别为1.223、0.398 g·mL^-1、214.803%、4.297%；优化后的浮性水产饲料微观表面相对光滑圆润，物料的熔融效果较好，结构质密，饲料膨化加工综合质量较好。