氧化乐果对雄性大鼠生殖毒性及作用机制

为评价氧化乐果在体内的亚慢性生殖毒性,以0（CK）、0.44、1.32和3.97mg/(kg•d).连续染毒大鼠6周。结果显示,氧化乐果能引起雄鼠体重下降,睾丸和附睾及脏器系数极显著增加（P ＜0.01）,睾丸组织病理学改变,尤其引起睾丸曲细精管萎缩、变性,各级生精细胞减少,支持细胞数量减少,间质增宽。降低附睾精子数及附睾精子活力(P ＜0.01 ）,精子畸形率显著升高(P ＜0.05 ）; 降低睾丸组织酸性磷酸酶（ACP）、碱性磷酸酶（AKP）、乳酸脱氢酶（LDH）的活力,增加γ-谷氨酰转移酶（γ-GT）活力（P ＜0.01）和睾酮（T）（P ＜0.01）和卵泡刺激素（FSH）的含量（P ＜0.05）。结合组织病理、性激素、酶活性及精子质量的变化,表明氧化乐果可对雄性大鼠机体产生明显的毒性作用,也具有雄性生殖毒性,其主要靶器官为睾丸。     

苹果短链脱氢酶基因MdSDR响应腐烂病菌侵染的功能研究

短链脱氢/还原酶(SDR)是一类NAD(P)(H)依赖的氧化还原酶,负责催化生物体内各种代谢活动的氧化还原步骤。为了探究苹果(Malus domestica)SDR(MdSDR)蛋白的功能,利用农杆菌介导的苹果组培苗瞬时转化技术在苹果叶片中瞬时表达MdSDR,然后通过刺伤接种法研究过表达MdSDR对叶片抗病性的影响;利用实时荧光定量PCR(qRT-PCR)技术检测瞬时表达MdSDR后苹果中脂肪酸生物合成相关基因的表达水平;利用气相色谱分析技术检测瞬时表达MdSDR苹果叶片中的脂肪酸含量;利用尼罗红染色检测苹果叶片中的脂滴(LDs)积累情况。结果表明,瞬时表达MdSDR显著增强了苹果叶片对腐烂病菌(Valsa mali)的抗病性,试验组叶片病斑直径相比对照组减小约14.46%(P<0.001);瞬时表达MdSDR后苹果叶片中脂肪酸生物合成相关基因的表达显著上调(P<0.001),其中MdACCase上调48.41倍,MdKAR上调129.79倍,MdENR上调168.20倍,MdHAD上调8.67倍,MdβCT、MdKASⅠ和MdKASⅡ均上调约5倍;然而过表达MdSDR后苹果叶片中脂肪酸的积累水平无显著变化(P>0.05);进一步研究发现,过表达MdSDR后苹果叶片中调控脂滴合成的基因MdSEIPIN显著上调表达(P<0.05),脂滴数量大量增加。本研究初步证明MdSDR能够通过促进脂肪酸的合成,间接提高脂滴的数量,从而提高苹果的抗病性,为苹果抗性品种的研究提供了一定的理论基础。     

过表达GmAP1.2转基因大豆的遗传转化及植株发育特性

为研究大豆GmAP1.2(Glyma.08G269800)的生物学功能,通过农杆菌介导的大豆子叶节遗传转化法将GmAP1.2基因遗传转化至大豆天隆1号,获得过表达GmAP1.2转基因植株;并对转基因大豆的株高、花器官、叶片等特性进行分析。结果表明,GmAP1.2过表达使转基因大豆比野生型(WT)早花31d,株高明显降低,且与野生型相比,GmAP1.2转基因大豆叶片变小,叶绿素a、叶绿素b和总叶绿素含量分别增加了1.43、1.52和1.57倍,但未影响花器官结构。表明GmAP1.2不仅能够影响大豆的株型,还能影响叶片的大小和叶绿素的含量。本研究结果为进一步阐明大豆AP1基因在大豆中的功能与应用奠定了理论基础。     

陕北典型农地表层土壤物理性质季节变化

土壤物理性质是影响土壤水文、侵蚀过程的重要因素。为此系统研究了陕北安塞4种典型农地（玉米、谷子、大豆和土豆）的土壤含水量、容重、水稳性团聚体和粘结力的季节变化特征。结果表明:4种农地表层土壤物理性质随季节变化的趋势大致相同,土壤含水量随降雨量的变化呈现出明显的峰谷。土壤容重和土壤粘结力在农作物生长阶段大体呈上升趋势,翻耕和收获后显著下降。水稳性团聚体则呈现单调递增趋势。在0.05显著性水平下的配对样本T检验显示:土壤含水量除玉米地和谷子地外（P=0.04）,其他农地土壤含水量之间无显著差异（0.29≤P≤0.99）;土壤容重除谷子地和大豆地外（P=0.03）,其他农地之间无显著差异（0.07≤P≤0.86）;土豆地与其他农地的水稳性团聚体之间有显著性差异（0.01≤P≤0.03）;各农地土壤粘结力之间无显著性差异（0.16≤P≤0.53）。土壤含水量的变化趋势与降雨的变化趋势基本一致,农事活动所引起的土壤扰动是土壤容重和粘结力变化的主要原因。研究结果对于分析农地水文、侵蚀过程的季节变化特征具有重要的意义。     

大豆鼓粒期叶片荧光参数与叶绿素含量的关系

叶绿素含量是研究大豆光合作用的重要生理指标,对大豆产量和品质具有重要影响,因此,进行叶绿素含量和光合功能的同步分析具有重要意义。本研究以鼓粒期大豆叶片为研究对象,对63个样本的叶绿素含量和荧光参数进行相关分析,建立回归模型,并用28个样本验证集进行验证及评价。结果表明,可变荧光与初始荧光比(F_v/F_o)、最大光化学效率(φ_PO)、初始时间点活性反应中心捕获的单个激子驱动除QA外的电子传递的效率(ψ_EO)、初始时间点用于电子传递的量子产额(φ_EO)、t=t_FM时单位面积内电子传递的量子产额(ET_O/CS_M)、性能指数(PI_ABS)、J点相对可变荧光(V_J)7个荧光参数与叶绿素含量相关性较好,相关系数分别为0.78、0.76、0.75、0.80、0.82、0.77、-0.75。回归模型方程为y=-0.138x₁+ 2.154x₂+0.002x₃+0.077x₄+0.076(R²=0.694)(x₁、x₂、x₃、x₄和y分别为F_v/F_o、φ_PO、ET_O/CS_M、 PI_ABS和叶绿素含量),验证模型决定系数(R²)=0.805 8,均方根误差(RMSE)=0.293 4,预测残差(RPD)=1.773 8,该模型具有较好的预测效果,可以丰富非生物逆境胁迫下无损监测大豆叶绿素估算方法。回归和通径分析发现,ET_O/CS_M对叶绿素含量直接作用最大,φ_PO次之,F_v/F_o对叶绿素含量起直接负作用,PI_ABS直接作用最小,直接通径系数分别为0.706、0.382、-0.303、0.078。本研究实现了大豆叶绿素含量与光合功能的同步分析,明确了荧光参数对叶绿素含量的影响效应,可为调节栽培措施,实现大豆高产提供理论基础。     

亚洲棉NCED3基因克隆及其抗旱功能分析

9-顺式-环氧类胡萝卜素双加氧酶(NCED)是脱落酸(ABA)合成过程中的关键限速酶,被证实广泛参与植物的生长发育进程及对非生物胁迫的应答。为了挖掘、鉴定棉花抗旱相关的NCEDs基因,本研究克隆了亚洲棉(Gossypium arboreum)GaNCED3基因,利用实时荧光定量PCR(qRT-PCR)技术分析了干旱胁迫下该基因的表达特性;并遗传转化拟南芥(Arabidopsis thaliana),研究了该基因在干旱应答中的功能。结果显示,GaNCED3基因开放阅读框(ORF)全长为1 794 bp,其编码的蛋白质包含597个氨基酸。该基因在干旱胁迫处理后上调表达,在处理3 h的根中表达量最高,是对照的27.6倍。在萌发期进行甘露醇模拟的干旱处理,超表达GaNCED3的转基因拟南芥种子萌发率和绿苗率均高于野生型。在幼苗期进行自然干旱处理后,转基因拟南芥植株叶片丙二醛含量显著低于对照(P<0.05),游离脯氨酸积累量显著高于对照(P<0.05)。本研究初步证明了外源超表达GaNCED3基因使植株抗旱能力增强,为进一步研究GaNCED3干旱应答的分子机制提供了理论依据,为抗旱育种提供了候选基因资源。     

草鱼Rab1A基因的克隆表达、抗体制备及对微囊藻毒素-LR的响应

为了解草鱼(Ctenopharygodon idella)小分子三磷酸鸟苷(GTP)结合蛋白家族成员Rab1A的分子结构特点及其在微囊藻毒素-LR (MC-LR)胁迫中的作用,本研究根据草鱼肝脏(MC-LR诱导)转录组测序获得的unigenes序列,采用RACE技术克隆了草鱼Rab1A基因cDNA,其全长序列为806 bp,开放阅读框为609 bp,编码202个氨基酸。同源性分析结果表明,Rab1A与鲤鱼(Cyprinus carpio) Rab1A3相似性最高。系统进化分析表明草鱼Rab1A与鲤鱼、斑马鱼(Danio rerio)等鱼类聚为一大支。实时荧光定量PCR (qRT-PCR)分析发现Rab1A在草鱼各组织中广泛分布,在血液和鳃等组织中的表达较丰富。构建了pGEX-4T-1-Rab1A重组表达载体,并制备了Rab1A多克隆抗体,通过酶联免疫(ELISA)检测抗体效价为1∶512 000以上,Western blot检测显示抗体具有特异性。草鱼肝脏经微囊藻毒素-LR(MC-LR)染毒,发现25和75 μg·g^-1感染96 h后对Rab1A基因和蛋白表达的影响不明显(P>0.05),但100 μg ·kg^-1 可导致草鱼肝脏Rab1A基因和蛋白表达显著下降 (P<0.05),表明Rab1A在参与草鱼抵御MC-LR胁迫过程中起着重要的作用。本研究为进一步了解Rab1A基因的功能及其在MC-LR胁迫过程中所发挥的作用奠定了基础。     

60Co-γ辐照模式下玉米中赤霉烯酮和品质的变化及对绵羊定时人工授精的影响

为研究经赤霉烯酮(ZEN)自然污染与⁶⁰Co-γ辐照处理后的玉米对绵羊定时人工授精中澳洲白种公羊、湖羊母羊的生殖性能的影响,本试验选取含ZEN玉米样本,分别用0、3、5、8、10 kGy剂量的⁶⁰Co-γ辐照,并测定辐照后玉米主要营养成分变化。然后分别将未污染玉米(Ⅰ对照组)、经10 kGy ⁶⁰Co-γ辐照的污染玉米(试验Ⅱ组)、污染玉米(试验Ⅲ组)饲喂定时人工授精的绵羊,检测澳洲白种公羊的精液质量,测定湖羊母羊的繁殖性能指标。结果表明,未经⁶⁰Co-γ辐照的玉米含ZEN 2 153 μg·kg^-1,在10 kGy ⁶⁰Co-γ辐照剂量下,玉米中ZEN降解率为83.7%;当⁶⁰Co-γ辐照剂量低于10 kGy时,玉米淀粉表面结构无变化,主要营养成分无显著变化。试验Ⅱ组澳洲白种公羊的采精量、精子活力、精子密度与试验Ⅲ组相比显著升高(P<0.05),与Ⅰ对照组相比无显著差异(P>0.05);试验Ⅲ组澳洲白种公羊第30天精子畸形率与试验Ⅱ组、Ⅰ对照组相比均显著升高(P<0.05),显微镜下观察可见双尾精子、双头精子、无尾精子等现象,其中双尾精子头部、尾部不完整;试验Ⅱ组湖羊母羊同期发情率、受胎率、产羔率和羔羊始重等指标与试验Ⅲ组相比显著升高(P<0.05),与Ⅰ对照组无显著差异(P>0.05)。综上,使用经10 kGy ⁶⁰Co-γ辐照后ZEN含量低于2 153 μg·kg^-1的玉米饲喂繁育绵羊,对绵羊定时人工授精无不利影响。本研究结果为⁶⁰Co-γ辐照模式下玉米中赤霉烯酮和品质的变化对绵羊定时人工授精的影响研究提供了理论依据。     

转Cry1Ac/Sck基因大米的免疫毒理学评价

为观察转Cry1Ac/Sck基因大米对小鼠免疫功能的影响,将BALB/c小鼠按体重分为4组,即转基因大米组、非转基因大米对照组、AIN93G饲料正常对照组和阳性对照组,相应饲料喂养30d,比较各组小鼠免疫功能改变情况。结果显示,转基因大米组血小板数量显著低于非转基因大米对照组,但是与正常对照组无显著性差异,并且这3组血小板数量均在正常变化范围内;其他观察指标转基因大米组与非转基因大米组相比均无显著性差异;阳性对照组的各项指标,绝大多数均显著低于其他3组。表明转Cry1Ac/Sck基因大米对小鼠免疫功能的影响与非转基因亲本大米基本相同,未对小鼠免疫功能产生不良影响,因此转Cry1Ac/Sck基因大米在小鼠免疫毒理学方面的评价是安全的。     

海藻糖对转C4型PEPC水稻种子萌发耐旱性的影响

为揭示海藻糖参与高表达转玉米C₄型磷酸烯醇式丙酮酸羧化酶(PEPC)基因(C₄-PEPC)水稻(Oryza sativa)(简称PC)在干旱胁迫下种子萌发的生理机制,本研究以PC及其未转基因野生型受体Kitaake(简称WT)种子为材料,研究外施不同浓度海藻糖联合干旱处理下,其种子活力、萌发过程中可溶性糖和脯氨酸含量、α-淀粉酶活性,以及α-淀粉酶、PEPC、糖信号、部分海藻糖相关基因的表达。结果表明,干旱处理均显著抑制了两材料的发芽率,并且抑制了芽的生长;外施低浓度海藻糖可缓解干旱对种子萌发的抑制,与WT相比,PC对海藻糖更加敏感,当海藻糖浓度为0.5 mmol·L^-1时已表现出明显的缓解效应,且较10 mmol·L^-1海藻糖对WT的缓解效果更佳。外施0.5 mmol·L^-1海藻糖联合干旱处理可以维持水稻的种子活力、种子内渗透调节物质含量以及α-淀粉酶活性,尤其有益于PC。外施0.5 mmol·L^-1海藻糖通过上调PC内依赖钙信号的CBL1-OsSnRK3.1/3.23基因的表达,激活OsK1a-OsMYBS1/2-OsAmy3/8途径;同时诱导OsTPP1/7合成海藻糖,上调蔗糖和葡萄糖含量,进而增强糖代谢,维持种子萌发。此外,海藻糖也上调了SAPK8/9/10和C₄-PEPC的转录水平,使PC在芽期表现耐旱。本研究结果为改善直播稻田的发芽率和“C₄稻”的创制提供了新线索。     

转基因大豆GTS40-3-2转化事件特异性PCR检测

GTS40-3-2是抗草甘膦转基因大豆,为建立GTS40-3-2大豆转化体特异性PCR检测方法,本研究以GTS40-3-2标准品为实验材料,根据已公布转基因大豆GTS40-3-2基因与大豆基因组连接序列信息,利用Primer5.0软件设计了5对品系特异性引物,对每对引物进行了退火温度、特异性及扩增效率的PCR检测,结果显示,5对特异性引物均能够从GTS40-3-2中扩增出大小约279bp、238bp、470bp、490bp和257bp的预期产物,可用于特异性检测转基因大豆GTS40-3-2转化事件。以转基因大豆GTS40-3-2含量为5%、2%、1%、0.5%和0.1%的标准品进行PCR灵敏度检测,结果表明5对引物的检测灵敏度均能达到0.1%。通过荧光定量PCR对5对特异性引物的Ct值与溶解曲线比较,最后选择出RRS2引物对为转基因大豆GTS40-3-2品系特异性检测的最适引物。本文结果将为我国未来转基因生物产品成分检测提供科学合理的实验参考。     

Event-specific detection of seven genetically modified soybean and maizes using multiplex-PCR coupled with oligonucleotide microarray

With the increasing development of genetically modified organism (GMO) detection techniques, the polymerase chain reaction (PCR) technique has been the mainstay for GMO detection. An oligonucleotide microarray is a glass chip to the surface of which an array of oligonucleotides was fixed as spots, each containing numerous copies of a sequence-specific probe that is complementary to a gene of interest. So it is used to detect ten or more targets synchronously. In this research, an event-specific detection strategy based on the unique and specific integration junction sequences between the host plant genome DNA and the integrated gene is being developed for its high specificity using multiplex-PCR together with oligonucleotide microarray. A commercial GM soybean (GTS 40-3-2) and six GM maize events (MON810, MON863, Bt176, Bt11, GA21, and T25) were detected by this method. The results indicate that it is a suitable method for the identification of these GM soybean and maizes.     

Establishment and application of event-specific polymerase chain reaction methods for two genetically modified soybean events, A2704-12 and A5547-127

For implementation of the issued regulations and labeling policies for genetically modified organism (GMO) supervision, the polymerase chain reaction (PCR) method has been widely used due to its high specificity and sensitivity. In particular, use of the event-specific PCR method based on the flanking sequence of transgenes has become the primary trend. In this study, both qualitative and quantitative PCR methods were established on the basis of the 5' flanking sequence of transgenic soybean A2704-12 and the 3' flanking sequence of transgenic soybean A5547-127, respectively. In qualitative PCR assays, the limits of detection (LODs) were 10 copies of haploid soybean genomic DNA for both A2704-12 and A5547-127. In quantitative real-time PCR assays, the LODs were 5 copies of haploid soybean genomic DNA for both A2704-12 and A5547-127, and the limits of quantification (LOQs) were 10 copies for both. Low bias and acceptable SD and RSD values were also achieved in quantification of four blind samples using the developed real-time PCR assays. In addition, the developed PCR assays for the two transgenic soybean events were used for routine analysis of soybean samples imported to Shanghai in a 6 month period from October 2010 to March 2011. A total of 27 lots of soybean from the United States and Argentina were analyzed: 8 lots from the Unites States were found to have the GM soybean A2704-12 event, and the GM contents were <1.5% in all eight analyzed lots. On the contrary, no GM soybean A5547-127 content was found in any of the eight lots. These results demonstrated that the established event-specific qualitative and quantitative PCR methods could be used effectively in routine identification and quantification of GM soybeans A2704-12 and A5547-127 and their derived products.     

Detection of transgenic and endogenous plant DNA fragments in the blood, tissues, and digesta of broilers

The aim was to determine the fate of transgenic and endogenous plant DNA fragments in the blood, tissues, and digesta of broilers. Male broiler chicks (n = 24) were allocated at 1 day old to each of four treatment diets designated T1-T4. T1 and T2 contained the near isogenic nongenetically modified (GM) maize grain, whereas T3 and T4 contained GM maize grain [cry1a(b) gene]; T1 and T3 also contained the near isogenic non-GM soybean meal, whereas T2 and T4 contained GM soybean meal (cp4epsps gene). Four days prior to slaughter at 39-42 days old, 50% of the broilers on T2-T4 had the source(s) of GM ingredients replaced by their non-GM counterparts. Detection of specific DNA sequences in feed, tissue, and digesta samples was completed by polymerase chain reaction analysis. Seven primer pairs were used to amplify fragments ( approximately 200 bp) from single copy genes (maize high mobility protein, soya lectin, and transgenes in the GM feeds) and multicopy genes (poultry mitochondrial cytochrome b, maize, and soya rubisco). There was no effect of treatment on the measured growth performance parameters. Except for a single detection of lectin (nontransgenic single copy gene; unsubstantiated) in the extracted DNA from one bursa tissue sample, there was no positive detection of any endogenous or transgenic single copy genes in either blood or tissue DNA samples. However, the multicopy rubisco gene was detected in a proportion of samples from all tissue types (23% of total across all tissues studied) and in low numbers in blood. Feed-derived DNA was found to survive complete degradation up to the large intestine. Transgenic DNA was detected in gizzard digesta but not in intestinal digesta 96 h after the last feeding of treatment diets containing a source of GM maize and/or soybean meal.     

甘南牦牛三种血液原虫多重PCR检测方法建立及流行病学调查

为建立一种能同时快速检测弓形虫、环形泰勒虫、新孢子虫3种甘南牦牛血液原虫的多重PCR检测方法,掌握3种血液原虫在甘南牦牛中的流行情况,本研究以弓形虫hypothetical protein基因、环形泰勒虫Tams1基因、新孢子虫Nc-p43基因为靶标设计3对特异性引物,通过优化多重PCR反应条件,建立检测3种血液原虫的多重PCR方法,并检测甘南地区629份牦牛血清样品,进行流行病学分析。结果显示,建立的牦牛3种血液原虫多重PCR检测方法,具有特异性强、敏感性高和重复性好的优点,检测弓形虫、环形泰勒虫和新孢子虫DNA的最低浓度分别为0.01、0.02、0.01 ng·μL-1;流行病学调查结果显示,弓形虫、环形泰勒虫和新孢子虫的阳性率分别为4.29%、3.66%、5.88%,并存在混合感染现象,整体混合感染率为13.68%;季节性流行病学分析显示,3种原虫单一感染率均为冬季最高、春季最低（P>0.05）,整体感染率为冬季最高、夏季最低（P>0.05）。按不同年龄分析,0～3岁牦牛3种原虫的整体感染率为27.83%,大于3岁的整体感染率27.46%(P>0.05)。分析风...     

麦冬、大黄饮片电子束辐照灭菌工艺研究

为了研究电子束辐照处理对麦冬、大黄饮片灭菌的工艺及效果,以麦冬、大黄饮片为材料,研究不同剂量(0、2、4、6、8 kGy)电子束辐照对其微生物总数、理化指标、指纹图谱相似度及有效成分含量的影响。根据不同堆码厚度、辐照方式计算剂量不均匀度,优化麦冬、大黄饮片的电子束辐照加工工艺。结果表明,电子束辐照能明显降低麦冬、大黄饮片中的微生物数量;电子束辐照对麦冬饮片水分、水溶性浸出物及总灰分含量的变化无显著影响(P>0.05);电子束辐照有利于提高大黄饮片水溶性浸出物含量,对其干燥失重、总灰分变化无显著影响(P>0.05);辐照前后麦冬、大黄饮片的指纹图谱相似度均大于0.99,相对保留时间(RSD)均小于0.5%;不同剂量电子束辐照对麦冬、大黄饮片有效成分含量均无显著影响(P>0.05);剂量不均匀度与产品堆码厚度及辐照方式密切相关。综上,麦冬饮片适宜辐照剂量为2~8 kGy,大黄饮片适宜剂量为4~8 kGy,此剂量范围对麦冬、大黄饮片整体质量无显著影响。单面辐照时,麦冬饮片适宜堆码厚度为4 cm,大黄饮片适宜堆码厚度为4~6 cm;双面辐照时,麦冬饮片适宜堆码厚度为10 cm,大黄饮片适宜堆码厚度为14 cm。本研究为麦冬、大黄加工贮藏及电子束辐照在中药中的应用提供了理论依据与技术支持。     

基于谱效分析评价电子束辐照灭菌对山银花药材质量的影响

为评价电子束辐照灭菌对山银花药材质量的影响,分别采用0、2.8、6.3、9.5、18.8 kGy吸收剂量的电子束辐照处理山银花及其提取物,研究不同剂量电子束辐照对其生物负载及指标成分绿原酸含量、抗氧化活性及指纹图谱的影响。结果表明,电子束辐照能够有效降低山银花及其提取物中需氧菌、霉菌和酵母菌总数,随着吸收剂量增大,其存活数显著降低;电子束辐照对山银花及其提取物中绿原酸的含量及1,1-二苯基-2-苦基肼自由基(DPPH·)清除率的影响不大,其变化与吸收剂量均无显著线性相关性(P>0.05),当吸收剂量达到18.8 kGy时,与未辐照样品相比均无显著差异(P>0.05)。不同吸收剂量电子束辐照处理的山银花及其提取物指纹图谱的相似度均为1;聚类分析(CA)结果也表明经不同吸收剂量电子束辐照处理的样品与未辐照样品未呈现显著差异。综上,电子束辐照灭菌对中药材山银花及其提取物的质量无明显影响。本研究利用谱效分析,系统评价电子束辐照灭菌对中药材山银花及其提取物质量的影响,为电子束辐照技术在山银花药材贮藏中的科学应用提供了参考依据。     

Changes in some soil properties in a Vertic Argiudoll under short-term conservation tillage

The purpose of this work was to determine whether some soil physical and chemical properties, and microbial activity were affected by two conservation tillage systems in a Chernozemic clay loam soil (Vertic Argiudoll), after 5 years of trial initiation. Two crop sequences, corn (Zea mays L.)–wheat (Triticum aestivum L.)/soybean (Glycine max (L.) Merr.) and wheat/soybean, under chisel plowing (ChP) and no till (NT) were evaluated. Physical and chemical properties were also analyzed taking the same soil without disturbance as reference. The Hénin instability index (HI) was larger in ChP than in NT in both corn–wheat/soybean (C–W/S) and wheat/soybean (W/S) sequences (P≤0.05). The C–W/S sequence differed from W/S (P≤0.01) in total organic carbon (TOC). As regards organic carbon fractions, no differences were found in labile organic carbon (LOC), while W/S under ChP showed the lowest value (P≤0.01) of humified organic carbon (HOC). No differences were found in microbial respiration either in crop sequences or in tillage systems. Soil physical and chemical properties differentiated crop sequences and tillage treatments from the undisturbed soil when a Student’s t-test was performed. Five years elapsed since the beginning of this trial was time enough to detect changes in some of the soil properties as a consequence of management practices. An important reduction in the soil structural stability was observed as related to the undisturbed soil. However, the C–W/S sequence under NT resulted in lower soil degradation with respect to the other treatments.