多溴联苯醚对大鼠肝细胞CYP1A2依赖性MROD活性的影响

采用未成熟SD大鼠肝细胞微粒体为试验材料,以2,3,7,8-四氯-二苯基-并-二哑噁英(2,3,7,8- tetrachlorobenzo-p-dioxin,TCDD)作为参照,利用甲氧基-异酚噁脱甲基酶(methoxyresorufin-O- demethylase,MROD)竞争性抑制动力学和荧光光谱法分析法,测定了5种多溴联苯(polybrominated diphenyl ethers,PBDEs)对7-甲氧基-异酚噁唑脱甲基反应的影响,间接研究了这些PBDEs对CYP1A2 的竞争性抑制作用。结果表明,和TCDD相比,PBDEs对MROD活性的抑制作用较弱,亦即对CYP1A2的 竞争性抑制作用较弱。     

固相微萃取-气相色谱-质谱联用法分析香蕉球茎象甲虫体挥发物

摘 要：为了探明香蕉球茎象甲虫体挥发物的各种成分,采用固相微萃取技术（SPME）提取香蕉球茎象甲的虫体挥发物,进行气质联用仪（GC-MS）分析鉴定。结果表明：香蕉球茎象甲雌虫、雄虫、雌雄虫（雌虫和雄虫等数量混合）分别鉴定出9种、8种和9种成分,各占总峰面积的61.05%,39.46%和17.43%。香蕉球茎象甲雌虫、雄虫和雌雄虫三种虫体挥发物共有成分有3,7-二甲基-1,3,6-辛三烯、α-荜澄茄油烯、(1,1-二甲基乙基)-2-甲氧基苯酚、邻苯二甲酸二乙酯、3,3',5,5'-四甲基联苯和邻苯二甲酸二异丁酯6种,而3,7-二甲基-1,3,6-辛三烯是香蕉球茎象甲雌虫、雄虫和雌雄虫三种虫体挥发物中含量最高的成分,为19.15%。香蕉球茎象甲虫体挥发物各成分种类多样,相对含量差异较大。     

玉米地方品种沃日黄与其选系重组改良后代表型性状综合分析

为评价玉米地方品种与其选系重组再选系方法的效率，探索优异地方种质资源发掘、利用新途径，采用表型分析、主成分分析、相关性分析与聚类分析，对‘沃30’与8个重组系进行综合评价，研究地方品种选系材料的综合评价指标和方法。结果表明：（1）‘沃30’与重组系间存在丰富的遗传变异，各性状变异系数为1.49%-46.79%，‘A3’和‘A4’的多个性状优于‘沃30’；（2）综合评价分析表明，3个主成分累计贡献率为90.46%，供试材料表型性状综合得分为-1.414-2.550，‘A4’、‘A3’和‘A1’综合得分高于‘沃30’，散粉期等6个性状与综合得分显著或极显著相关；（3）供试材料间遗传距离为0.78-2.045，当阈值为1.414，可分为三大类。通过地方品种‘沃日黄’优株与其选系重组再选系的方法能够改良目标性状，利于进一步发掘与利用地方品种中的优良基因。     

不同品种水稻对乙草胺的敏感性研究

通过测定乙草胺对16个品种水稻生长的影响,研究不同品种水稻对乙草胺的敏感性及其与品种类型之间的关系。培养皿法测定乙草胺对水稻发芽的影响,琼脂法测定乙草胺对各品种水稻苗、根长度及重量等指标的抑制作用。乙草胺对水稻种子发芽有抑制作用;低浓度时乙草胺对水稻的株高、根数有促进作用,当乙草胺浓度达到一定值,则对水稻生长有较强的抑制作用,且对根的抑制作用比对茎的抑制作用更强;供试品种中‘泗稻’耐药性最强,‘天优3301’敏感性最高,0.02 mg/L乙草胺琼脂培养基培养对2个水稻品种的苗长、苗鲜重、根长、株鲜重及根数平均抑制率分别为1.46%、6.42%、51.06%、60.87%和-6.45%（‘泗稻’）;21.62%、22.62%、63.16%、74.76%、3.89%（‘天优3301’）。不同品种水稻对乙草胺的敏感性存在显著差异,且该差异与品种类型相关性不强。     

速眠新对大鼠原代肝细胞CYP3A1、CYP2E1的影响

胶原酶二步灌流法获取原代大鼠肝细胞,用不同浓度的速眠新和保定宁处理,Western blot法测定肝细胞中细胞色素酶P450 3A1（CYP3A1）和细胞色素酶P4502E1 (CYP 2E1)的表达水平。结果表明：通过胶原酶灌流法,每只大鼠可获得2-4×108个肝细胞,成活率约为95%。用速眠新、保定宁处理后,肝细胞CYP3A1和CYP 2E1的表达随速眠新、保定宁浓度的增加和处理时间的延长而呈升高的趋势。原代大鼠肝细胞可用于速眠新药物代谢分子机制的研究,为其临床上合理地应用提供了理论依据。     

3个烤烟品种（系）在安顺烟区适应性的研究

为了优选适应安顺特色优质烟叶开发的品种,以‘2010-6’、‘A18’、‘贵烟2号’3个烤烟品种（系）为参试品种,以‘K326’为对照在安顺西秀区进行生产试验,分别从生长状况、农艺性状、抗病性、产量及烟叶评吸质量等方面对各参试品种（系）及对照进行分析比较,鉴定参试品种（系）对当地土壤、栽培、烘烤条件的反应和再生产价值。试验结果表明：各参试品种（系）在安顺市西秀区的综合表现来看,均好于对照品种‘K326’,其中以‘贵烟2号’表现最好,‘2010-6’、‘A18’略优于对照品种‘K326’。3个参试品种（系）可作为安顺市特色优质补充品种进一步试验研究。     

引进番茄品种抗晚疫病苗期鉴定及抗性品种筛选

以俄罗斯番茄品种A5、A6、A8、A9、A10、7-1、20、富贵（中国）和日本品种耐运2000为试材进行番茄苗期抗晚疫病鉴定及抗性品种筛选,研究表明：（1）最适的孢子囊悬浮液接种浓度为1×103孢子囊/ml;（2）同一品种幼苗比离体叶片病情指数普遍偏低;（3）耐运2000 、A10抗病性最强（抗病品种）,A9、20、7-1、A5和A8中等（感病品种）,富贵和A6最差,属于高感品种。因此耐运2000和A10有希望成为抗晚疫病品种或作为进一步培育抗晚疫病品种的亲本。     

1-甲基环丙烯结合气调处理对‘帕拉英达’和‘吉禄’芒果保鲜效果的影响

为探究1-甲基环丙烯（1-Methylcyclopropene, 1-MCP）结合气调处理对采后‘帕拉英达’和‘吉禄’芒果贮藏品质的影响,以元江晚熟芒果品种‘帕拉英达’和‘吉禄’为试材,分别用0.5 μL/L 1-MCP、微孔膜、PBI气调保鲜袋、1-MCP+微孔膜及1-MCP+PBI气调保鲜袋对芒果进行处理,对其外观指标、失重率、硬度、可溶性固形物（TSS）含量、VC含量、可滴定酸（TA）含量、呼吸强度、商品率以及呼吸代谢相关酶活性变化进行检测分析。结果表明：在（13±1）℃贮藏过程中,与未作任何处理的对照组相比,5种处理在一定时间内均能延缓‘帕拉英达’和‘吉禄’果实黄化指数、病情指数、腐烂指数、色差、失重率、可溶性固形物含量和呼吸强度的上升,抑制果实硬度、VC含量、可滴定酸含量及商品率的下降,显著抑制果实呼吸代谢相关酶活性变化。综合分析,在（13±1）℃贮藏环境中,1-MCP+PBI气调保鲜袋处理对‘帕拉英达’和‘吉禄’芒果的保鲜效果最优,更有利于果实的贮藏。研究结果可为低纬度、高海拔地区晚熟芒果采后保鲜提供技术参考和科学依据。     

不同基因型烤烟对镰刀菌根腐病的抗性鉴定与评价

为明确不同基因型烤烟对烟草镰刀菌根腐病的抗性,利用10个烟草品种（系）,采用粗毒素幼芽接种、带菌菌谷接种、孢子悬浮液培养等方法进行抗性鉴定。结果表明,供试品种对尖孢镰刀菌抗性表现出显著差异;对尖孢镰刀菌表现出抗的品种（系）有4个,分别是‘Y2001’、‘Y2002’、‘长脖黄’、‘664-01’;对尖孢镰刀菌表现出感的品种（系）有6个,分别是‘7710’‘、7713’‘、Y2007’‘、Y2008’‘、中烟100’、‘小黄金1025’。3种接种方法鉴定结果基本一致,共筛选出4个抗性品种（系）。     

甘蔗与蔗茅杂交亲本及后代材料的抗旱性鉴定

研究旨在筛选出优良抗旱育种材料,为今后甘蔗抗旱育种和抗旱栽培技术提供重要的理论依据。采用盆栽干旱处理,测定蔗茅野生种无性系、甘蔗栽培品种（系）及其杂交后代共12份材料苗期与抗旱相关的8项生理生化指标,利用极点排序法对供试材料的抗旱性进行了综合评价。结果表明：干旱胁迫下,12份供试材料的Pro、MDA、SS及PMP均呈上升趋势,SP、Chl和SOD活性呈下降趋势,CAT活性有升有降。极点排序法分析表明,12个供试材料的抗旱强弱为：‘蔗茅99-2’>‘蔗茅99-1’>‘蔗茅99-4’>‘蔗茅I91-8’>‘蔗茅99-3’>‘蔗茅90-29’>‘滇蔗01-58’>‘滇蔗04-14’>‘滇蔗02-39’>‘蔗茅II91-2’>‘新台糖10’>‘崖城89-9’,说明蔗茅优质抗旱性状可以通过基因渗透方式传递给后代。在苗期干旱胁迫下,‘蔗茅99-2’在渗透调节等方面表现出积极的生理响应。     

Analysis of Cotton Secondary Metabolites under the Interaction between Verticillium dahliae and Upland Cotton

[Objective] The aim of this study is to investigate the cotton differential metabolites in the interaction between Verticillium dahliae and cotton plant, and to explore new clues for the further study on the defense mechanism of cotton against V. dahliae. [Method] To obtain the cotton root, stem and leaf samples of the pathogen treated and healthy controls, the upland cotton cultivar CCRI 24 was selected as tested cotton material in this paper, and inoculated with V. dahliae conidia or sterile water by injuring root at the 2-true-leaf stage. The 70% (volume fraction) methanol extracts of these samples were separated and detected by ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS), and the metabolite data were obtained by the online XCMS software. Through multivariate statistics and student’s t-test, the differential metabolites were investigated. The types of metabolites were putatively identified based on the comparison of the experimental molecular mass and the monoisotopic accurate molecular mass of cotton metabolites. [Result] The efficiency of negative ion mode was higher than that of positive ion detection mode in the cotton metabolites in UPLC-ESI-MS analysis. 576 ions mainly distributed in cotton root were found in the cotton seedling tissues including root, stem and leaf, which were the distinguished differential metabolites between V. dahliae treatment and healthy control. Among them, 77 ions were identified as sesquiterpenoids, diterpenoids, flavonoids, carbohydrates, aliphatics and phenols. In addition to sesquiterpenes, 17 compounds, including caffeic acid, astragalin, isoastragalin, dillenetin, ent-catechin, gossypetin 8-rhamnoside, gossypicyanin, herbacetin 7-glucoside, leucocyanidin, quercetin 3'-glucoside, quercetin 3-glucoside, quercetin 7-glycosides, α,2',3,3',4,4',6-heptahydroxychalcone 2'-glucoside, melibiose, sucrose, sucrose 6-phosphate and 1-tetratriacontanol, had not been reported in the literatures on the interaction between V. dahliae and cotton, which may be the novel pathogenesis-related metabolites on cotton Verticillium wilt. [Conclusion] The putative pathogenesis-related metabolites of cotton Verticillium wilt may play an important role in the interaction of cotton defense against V. dahliae, which provides an important clue for exploring the new resistant mechanism of cotton to Verticillium wilt.     

绞股蓝皂苷生物合成后修饰酶基因的cDNA克隆和组织表达特征

从绞股蓝转录组挖掘可能与绞股蓝皂苷生物合成相关修饰酶基因细胞色素P450(cytochrome P450,CYP)及UDP-糖基转移酶(UDP-dependent glycosyltransferase,UGT)基因的cDNA进行克隆并开展了以下研究。根据课题组前期绞股蓝转录组数据,筛选CYP及UGT Unigenes,利用RT-PCR克隆ORF全长,并结合生物信息学对其编码蛋白进行功能分析,最后通过荧光定量PCR验证其在根、茎、叶的差异性表达。结果克隆得到两个ORF序列,分别命名为CYP94A1和UGT91A1;CYP94A1序列包含1509 bp的开放阅读框架,编码一条含503个氨基酸残基的肽链,具有CYP保守结构域;UGT91A1序列包含1383 bp的开放阅读框架,编码一条含461个氨基酸残基的肽链,具有UGT保守结构域。这两个基因的转录表达均具有组织特异性,在叶或茎中的表达均高于根。CYP94A1和UGT91A1的克隆、转录表达及生物信息学分析为进一步挖掘绞股蓝中CYPs、UGTs及功能验证提供了帮助,增加对CYP和UGT两个超基因家族的认识。     

樟叶越桔CYP450基因片段克隆与家族分析

广泛存在于生物体内的CYP450具有参与多种内源物质的新陈代谢和解除外源化学物质毒害作用的功能。本研究基于RNA-Seq数据,应用RT-PCR技术首次从樟叶越桔中克隆获得21个CYP450基因cDNA片段。应用生物信息学技术对21个樟叶越桔CYP450基因进行相似性分析、功能注释、家族分类以及聚类分析,除2个基因的家族分类不明确外,其余所有基因被分属于10个不同的CYP450家族中,分别为:CYP71、CYP72、CYP76、CYP81、CYP82、CYP86、CYP94、CYP97、CYP734和CYP735。推测有10个基因参与脂肪族、芳香族碳的羟基化,4个基因参与植物激素细胞分裂素的合成与降解,3个基因参与脂肪酸的羟基化,2个基因参与萜类化合物的合成。而KEGG分析结果显示2个基因家族未知的樟叶越桔CYP450基因可能参与了K00327途径。21个樟叶越桔CYP450基因能成功地与24个拟南芥CYP450基因聚类到一起。值得关注的是有2个樟叶越桔CYP450基因CL75.Contig12A(9)和Unigene225702A(14)被归类到拟南芥所特有的CYP82家族中,分别属于CYP82D和CYP82G亚家族,这为研究CYP82基因家族新物种来源提供了参考。本研究结果为认识樟叶越桔细胞色素CYP450基因和家族分类提供了信息,为樟叶越桔CYP450功能研究提供了参考依据。     

Inheritance of berry volatile compounds in two half-sib grape (Vitis vinifera) populations

Inheritance of grape berry volatile compounds was studied in the half-sib populations of ‘Jingxiu’ × ‘Muscat of Alexandria’ and ‘Jingxiu’ × ‘Xiangfei’ during two successive years. Twenty-two volatile compounds were identified via GC–MS. Monoterpenes were detected in the paternal parents ‘Muscat of Alexandria’ and ‘Xiangfei’, which were Muscat grapes, but not in the non-Muscat maternal parent ‘Jingxiu’. Four C6 compounds, five carbonyls and one ester (ethyl acetate) were found in all the parents, 2-ethylhexanol (alcohol) in ‘Jingxiu’ and ‘Muscat of Alexandria’, and hexyl acetate (ester) in ‘Jingxiu’. The paternal effect varied with year, while the inheritance patterns of volatile compounds, except for (Z)-linalool oxide, were stable between years and between populations. Four C6 compounds, five carbonyls, two esters and one alcohol were typical quantitative traits in both half-sib populations in 2 years. Most monoterpenes, including geraniol, α-terpineol and linalool, exhibited continuous variation in the half-sib progenies, but some other monoterpenes, such as (Z)-rose oxide, nerol oxide, nerol, neral, geranial and geranic acid, appeared to segregate into presence or absence in the progenies (1:1). However, (Z)-linalool oxide, behaved as a quantitative trait in the ‘Jingxiu’ × ‘Muscat of Alexandria’ population, but segregated into presence or absence (1:1) in the ‘Jingxiu’ × ‘Xiangfei’ population for both years.     

一个与氰苷生物合成相关的植物CYP基因的克隆与表达分析

氰苷是由植物细胞色素P450单加氧酶(CYP)催化氨基酸生成的次生代谢产物,与植物的防御和抗逆境胁迫相关。为了获得蒜头果中与氰苷生物合成有关的CYP基因,本研究通过分析蒜头果转录组数据,从中分离并克隆得到1条细胞色素P450基因(命名为MoCYP79),并对其进行生物信息学分析及检测该基因在果实不同发育时期的具体表达情况。结果显示,MoCYP79基因的cDNA全长1632 bp,编码543个氨基酸;MoCYP79蛋白含有CYP家族的识别结构域血红素结合域(FSTGRRGC),且与巨桉(XP_010027351.1)、木薯(AAF27290.1,AAF27289.1)等植物CYP79蛋白聚在一支;MoCYP79基因在蒜头果花谢后的果实中表达量呈下降趋势,除发育3个月的果实外,该基因在其余的果实中的表达量均显著高于叶片,从大到小依次为花谢后的果实1个月>2个月>4个月3个月。本实验对研究蒜头果的对病虫害的防御、逆境胁迫的抵御及蒜头果中有效次生代谢产物的发掘具有重要的意义。     

辣椒种子引发技术优化试验

本文对辣椒种子引发方法进行了优化。以不同品种的辣椒种子为试材,利用振荡器作引发工具,用不同化学药剂和H2O作引发剂,通过测定引发过的辣椒种子的发芽特性,筛选最佳引发剂配方和引发时间。实验结果表明,引发后的种子发芽特性和抗逆性均增强,处理（1）‘1.8% KNO3+0.2% ZnSO4,处理4天’的综合表现最佳,处理（4）‘10% PEG+1.8% KNO3+0.2% ZnSO4,处理6天’的种子在半个月内发芽特性最好,但3个月后发芽率明显下降。由此可见,处理（1）‘1.8% KNO3+0.2% ZnSO4,处理4天’为最佳引发方法。     

Effects of Foliar Application of Elicitors on Red Clover Isoflavone Content

Red clover (Trifolium pratense L.) contains high concentrations of isoflavones, compounds that have received much interest lately due to their presumed benefits for human health. In this experiment we tested the possibility to induce isoflavone production in the foliage of two greenhouse‐grown red clover cultivars (‘Azur’ and ‘Start’) through the application of elicitor compounds. Foliar applications of different concentrations of acetic acid (50, 100, 250 and 500 mm ), yeast extract (1, 2, 3 and 4 g l^?1), and chitosan (125, 250, 500, and 1000 mg l^?1) were carried out on plants at the late vegetative stage, which were harvested 2 or 8 days after spraying. Concentrations of genistein, daidzein, formononetin and biochanin A were determined by high performance liquid chromatography. The two cultivars differed in isoflavone concentrations, ‘Azur’ having on average 36 % higher biochanin A, formononetin and total isoflavone concentrations than ‘Start’ (P < 0.05). A cultivar × sampling date interaction (P < 0.1) reflected a 20 % increase over time in total isoflavone concentration with ‘Azur’, which was not observed with ‘Start’. Effects of elicitors were limited, contrasts indicating overall, 12, 14 and 15 % greater total isoflavone concentration in yeast extract (P < 0.1), chitosan (P < 0.05) and acetic acid (P < 0.05)‐treated plants, respectively, than in untreated control plants. There were few differences between the various elicitors and none between concentrations of each elicitor.     

3个杏品种果实香气顶空固相微萃取气质联用分析

对三个杏品种“金太阳”、“鲁杏1号”、“鲁杏2号”成熟期果实顶空微萃取果肉中的挥发性物质进行气相色谱—质谱分析鉴定。金太阳杏检测出33种香气成分,占总峰面积的82.37%,主要香气物质为乙酸己酯、乙酸－4－己烯－1－酯、二羟乙酸、6－甲基－5－烯基－2－庚酮;“鲁杏1号”检测出10种香气成分,占总峰面积的51.18%,主要香气物质为二羟乙酸、苯乙醇、二乙基邻苯二甲酸酯、2-苯并噻吩;“鲁杏2号”检出10种香气成分,占总峰面积的48.06%。主要香气物质为乙醇、乙酸己酯、顺-3-己烯基-乙酸酯。三个品种间香气成分和含量均存在差异,杏香气成分是杏选育评价标准的重要指标之一。