豆大蓟马对12种杀虫剂的敏感性测定

室内采用叶管药膜法测定12种杀虫剂对豆大蓟马(Megalurothrips usitatus)成虫和若虫的毒力。结果表明,供试药剂中除杀虫环和拟除虫菊酯类杀虫剂对豆大蓟马各虫态的毒力较低外,其他药剂均对豆大蓟马具有较高的毒力。毒力测定分析结果表明,供试药剂对豆大蓟马若虫的毒力不同程度上均大于成虫,且对成虫的毒力又以对雄成虫的毒力大于雌成虫。综合来看,乙基多杀菌素、甲维盐、噻虫胺、啶虫脒、阿维菌素和噻虫啉等药剂对豆大蓟马毒力较大,可推荐作为田间防治药剂。     

马铃薯黑痣病生防菌的筛选和鉴定及其生长条件的研究

从甘肃省景泰县条山农场不同连作年限的马铃薯地块中取根际土样品,分离筛选马铃薯黑痣病病原菌-立枯丝核菌(Rhizoctonia solani)的拮抗菌。采用土壤平板稀释、对峙法筛选拮抗菌;结合形态特征、生理生化及16Sr DNA分子鉴定;采用单因素试验设计对G1菌株的生长条件进行初步试验。通过初筛和复筛,从108株细菌中获得1株对立枯丝核菌拮抗效果稳定、拮抗能力较强的细菌G1。采用平板对峙法测定其拮抗作用,抑菌带宽度达6mm,抑菌圈直径2.5 cm,抑菌率为66.7%。结合形态特征、生理生化及16Sr DNA分子鉴定,G1为类芽孢杆菌(Paenibacillus sp.);采用单因素试验设计对G1菌株的生长条件进行初步试验。结果表明,菌株G1在4℃~45℃,p H 5~10,盐浓度0~50 g·L-1都能生长,碳源、氮源利用广泛,最适条件为:温度37℃,p H 7,蔗糖为碳源,蛋白胨为氮源,盐浓度为10 g·L-1。拮抗菌G1被鉴定为类芽孢杆菌,该菌株抑菌带较宽、抑菌效果稳定,生长条件与马铃薯生长的大田环境基本相符,具有作为一株优良生防菌的必要条件。     

Soil carbon storage and stratification under different tillage/residue-management practices in double rice cropping system

The importance of soil organic carbon(SOC) sequestration in agricultural soils as climate-change-mitigating strategy has become an area of focus by the scientific community in relation to soil management.This study was conducted to determine the temporal effect of different tillage systems and residue management on distribution, storage and stratification of SOC, and the yield of rice under double rice(Oryza sativa L.) cropping system in the southern China.A tillage experiment was conducted in the southern China during 2005–2011, including plow tillage with residue removed(PT0), plow tillage with residue retention(PT), rotary tillage with residue retention(RT), and no-till with residue retention on the surface(NT).The soil samples were obtained at the harvesting of late rice in October of 2005, 2007 and 2011.Multiple-year residue return application significantly increased rice yields for the two rice-cropping systems; yields of early and late rice were higher under RT than those under other tillage systems in both years in 2011.Compared with PT0, SOC stocks were increased in soil under NT at 0–5, 5–10, 10–20, and 20–30 cm depths by 33.8, 4.1, 6.6, and 53.3%, respectively, in 2011.SOC stocks under RT were higher than these under other tillage treatments at 0–30 cm depth.SOC stocks in soil under PT were higher than those under PT0 in the 0–5 and 20–30 cm soil layers.Therefore, crop residues played an important role in SOC management, and improvement of soil quality.In the 0–20 cm layer, the stratification ratio(SR) of SOC followed the order NTRTPTPT0; when the 0–30 cm layer was considered, NT also had the highest SR of SOC, but the SR of SOC under PT was higher than that under RT with a multiple-year tillage practice.Therefore, the notion that conservation tillage lead to higher SOC stocks and soil quality than plowed systems requires cautious scrutiny.Nevertheless, some benefits associated with RT system present a greater potential for its adoption in view of the multiple-year environmental sustainability under double rice cropping system in the southern China.     

雌激素和孕酮对犬子宫内膜基质细胞增殖和孕酮受体表达的影响

本试验应用不同消化分离途径获取犬子宫内膜基质细胞,调节培养液中雌激素(E₂)和孕酮(P₄)的浓度,采用MTT法测定E₂和P₄浓度水平对犬子宫内膜基质细胞体外增殖的影响,利用细胞免疫组织化学法鉴定细胞并测定细胞孕酮受体(PR)表达与激素浓度水平的相关性。结果表明,E₂浓度变化(15、30、100 pg/mL)对犬子宫内膜基质细胞的增殖和PR的表达均没有显著的调节作用(P>0.05);P₄(15、30 ng/mL)对犬子宫内膜基质细胞的增殖有显著促进作用(P<0.05),P₄(3、15、30 ng/mL)对犬子宫内膜基质细胞PR的表达具有显著的抑制作用(P<0.05),其影响程度与浓度和作用时间关系密切。     

多能性转录因子mRNA转染山羊胎儿成纤维细胞条件优化

【目的】针对转染试剂脂质体2000和转染方法进行研究,用以优化外源多能性转录因子oct4、sox2、klf4、c-myc的mRNA转入山羊胎儿成纤维细胞（goat embryonic fibroblast,GEF）的条件。【方法】采用胰酶消化法,从山羊胎儿腿部肌肉组织中分离并获得了纯化的GEF细胞。根据mMESSAGE mMACHINE® T7 Ultra Kit试剂盒说明书,体外转录步骤主要有：①前期的质粒准备：首先用质粒小提试剂盒对摇菌获得的各体外转录载体质粒进行提取,随后对各体外转录载体目的基因下游进行单酶切,获得线性化质粒,最后用DNA纯化试剂盒对线性化的质粒进行纯化;②mRNA的“加帽”：以线性化DNA为模板,合成带有5′端帽子结构的mRNA,添加TURBO DNase以除去模板DNA;③mRNA的“加尾”：以ATP为原料,利用试剂盒自带的Poly(A)聚合酶,对已经“加帽”的mRNA进行“加尾”,以合成结构完整的mRNA;④完整mRNA的纯化：按照MEGAclear^TM Kit试剂盒对mRNA进行纯化。随后,将体外转录获得各转录因子的mRNA进行浓度和OD值测定。分别比较总mRNA（EGFP和4种多能性转录因子的mRNA的质量分别为0.2μg）和脂质体2000 的比例为1:0.5、1:1.0、1:1.5、1:2.0以及在最佳总mRNA与脂质体2000比例体系下第2、4、6代GEF细胞的mRNA转染效率。对最佳转染体系下转染多能性转录因子mRNA的GEF细胞进行免疫荧光检测,并对转染多能性转录因子mRNA后GEF细胞的形态和数目进行观察。【结果】mRNA和脂质体2000 的比例为1:1.0以及在最佳总mRNA与脂质体2000比例体系下第2代GEF细胞的mRNA转染效率均显著高于其他组（P＜0.05）,且本组细胞在形态和生长方面相对较好;免疫荧光检测表明,4种多能性转录因子的mRNA在GEF细胞中定位于细胞核,并获得稳定表达,在细胞质中不见多能性转录因子oct4、sox2、klf4、c-myc的表达;GEF细胞在转染4种mRNA 24h后,细胞形态由梭状慢慢向圆形靠拢;细胞活力低于对照组（山羊成纤维细胞正常生长组）。【结论】GEF细胞在总mRNA和脂质体2000 的比例为1:1.0以及在最佳总mRNA与脂质体2000比例体系下第2代GEF的条件下更适合mRNA的转染。研究工作为mRNA转染体细胞或干细胞的研究及诱导性多能干细胞（induced pluripotent stem cells,iPS）的获得提供参考资料,并对mRNA在成纤维细胞中的去向提供间接证据,有利于更加深入地了解多能性转录因子oct4、sox2、klf4、c-myc的mRNA相关功能。     

Water in the Mogao Grottoes,China: where it comes from and how it is driven

The Dunhuang Mogao Grottoes in China was designated as a world heritage site by UNESCO in 1987 and is famous for its cultural relics. Water is the most active factor that harms the relics in the caves as it damages the grotto murals and painted sculptures. Thus, determining the water sources and driving forces of water movement is a key issue for protecting these cultural relics. These issues have troubled relics protectors for a long time. In this study, the authors chose a representative cave in the Mogao Grottoes and, by completely sealing the cave to make a closed system, measured the water vapor from the surrounding rock. This was accomplished by installing a condensation–dehumidification temperature–humidity control system for the collection of water vapor. The results show that there is continuous evaporation from the deep surrounding rock into the cave. The daily evaporation capacity is determined to be 1.02 g/(d?m2). The water sources and driving forces of water movement were further analyzed according to the character of the water evaporation and by monitoring the temperature and humidity of the surrounding rock. It was found that the water vapor in the cave derives from phreatic water. Moreover, the yearly fluctuation of temperature in the surrounding rock and geothermal forces are the basic powers responsible for driving phreatic evaporation. Under the action of the yearly temperature fluctuations, decomposition and combination of bound water acts as a "pump" that drives phreatic water migration and evaporation. When the temperature rises, bound water decomposes and evaporates; and when it falls, the rock absorbs moisture. This causes the phreatic water to move from deep regions to shallow ones. Determining the source and dynamic foundation of the water provides a firm scientific basis for protecting the valuable cultural relics in the caves.     

山东半岛南部海域三疣梭子蟹增殖放流群体回捕率

根据2010年和2011年5-8月在山东半岛南部海域对三疣梭子蟹(Portunus trituberculatus)增殖放流前后分别进行的每年3个航次的对比调查结果,定量分析了三疣梭子蟹自然群体和增殖放流群体的数量,并根据三疣梭子蟹放流苗种数量和放流后捕捞产量,对三疣梭子蟹增殖放流群体回捕率进行分析.研究结果表明,2010年5月山东半岛南部海域三疣梭子蟹增殖放流前自然群体资源量很少,相对资源密度为0.90 ind/(站.h);在5-6月共放流苗种11 015.13万只,放流后10d左右资源量增加为26.08 ind/(站.h),放流群体所占的比例为96.55％;8月进行调查的相对资源密度为1.70 ind/(站.h),当年生群体数量占所有群体数量的比例为64.29％;9月、10月、11月秋汛期间捕捞产量为3 108 t,捕捞个体平均质量为232.3 g,增殖放流群体回捕率为7.54％.201 1年5月山东半岛南部海域三疣梭子蟹增殖放流前自然群体相对资源密度为1.70 ind/(站.h);在5-6月共放流苗种13 132万只,放流后10d左右资源量增加为29.15 ind/(站.h),放流群体所占的比例为94.17％;8月进行调查的相对数量为1.00 ind/(站.h),当年生群体数量占所有群体数量的比例为70.00％;9月、10月、11月秋汛期间捕捞产量为2 896 t,回捕个体平均质量为226.0 g,增殖放流群体回捕率为6.43％.除2011年胶州湾海域由于富营养化水质较差,以及其他因素对黄家塘湾海域的影响导致这两个放流点放流群体回捕率偏低外,2010年和2011年山东半岛南部放流海域整体水质良好,水温、透明度、盐度、营养盐比例、生物饵料等均比较适中,属于放流环境条件较佳的海域,比较适合三疣梭子蟹生长,放流群体回捕率整体上处于较高水平.     

斑节对虾细胞周期蛋白B慢病毒载体的构建及对Hela细胞增殖影响

细胞周期蛋白B（Cyclin B）是影响卵母细胞发育成熟的重要基因。文章构建了PmCyB慢病毒表达载体,并和慢病毒包装复合物在293T细胞中包装成慢病毒颗粒。利用包装好的慢病毒颗粒,感染干扰了Cyclin B1基因的Hela细胞,研究了PmCyB对Hela细胞增殖的影响。结果表明,通过siRNA干扰细胞自身的Cyclin B1后,转染对照组病毒液的Hela细胞增殖减缓;同时,干扰后转入含有PmCyB重组慢病毒表达载体,Hela细胞增殖速度明显要高于转染对照组病毒液的Hela细胞的增殖速度。这一结果表明,PmCyB基因的过表达能补偿Cyclin B1缺失导致的细胞增殖减缓的现象,说明PmCyB基因具有细胞周期调控功能,是细胞增殖与分化的重要调控基因,且功能可能与人类的Cyclin B1相似。人类的Cyclin B1是卵母细胞发育成熟的重要基因。由此可以推断,PmCyB基因可能对斑节对虾（Penaeus monodon）卵巢发育成熟起着重要作用。     

添加糖蜜和乙酸对西藏发酵全混合日粮青贮发酵品质及有氧稳定性影响

为研究乙酸和糖蜜对发酵全混合日粮(FTMR)品质及有氧稳定性的影响,试验以全混合日粮(TMR)为发酵原料,设对照组、乙酸(A)、糖蜜(M)、乙酸和糖蜜组合添加(AM)4个处理,发酵45 d后分析FTMR的发酵品质,同时将所有FTMR暴露到空气中,分别在第6,9和12天取样评定其有氧稳定性。结果表明,与对照相比,A和AM组降低了乳酸含量(P>0.05),各组仅检测到微量的丙酸、少量的丁酸和较低的氨态氮/总氮,表明各组FTMR发酵品质均良好。有氧暴露6 d后,各组乳酸、乙酸和水溶性碳水化合物含量均呈不同程度的下降,对照和M组pH值在第12 天显著(P<0.05)上升至5.70和6.50,具有较高的氨态氮/总氮和酵母菌数量。而A和AM组乳酸含量均呈先上升后下降的变化趋势,且好氧性微生物和酵母菌数量在整个有氧暴露的过程中始终维持在较低水平,有氧暴露12 d后pH值仍维持在4.50左右,延长了有氧稳定时间。综合考虑,为了获得品质优良的FTMR饲料,可在TMR中添加0.3%乙酸,既不影响发酵品质,也可提高其有氧稳定性。     

芝麻枯萎病病原菌致病力室内鉴定方法

 芝麻枯萎病(Sesame Fusarium wilt, SFW)是由尖孢镰刀菌芝麻专化型(Fusarium oxysporum Schl. f. sp. sesami (Zap.), FOS)引起的一种土传真菌病害, 是世界芝麻生产上的主要病害之一。为测定FOS致病力, 本文选用郑芝98N09等4个芝麻品种, 在苗期对15个FOS菌株的致病力强弱进行了室内鉴定和评价。结果表明, 采用1×10⁶个/mL分生孢子悬浮液与无菌蛭石和无菌土壤按V1∶V2∶V6比例混合接菌(即最终接菌浓度为1.4×10⁵孢子/g土壤), 在接菌后第7 d幼苗开始出现枯萎病症状, 调查菌株致病性的最佳时间为接菌后第25 d~28 d;在供试15个FOS菌株中, 对4个品种均表现为强致病力的菌株有8个(DI>50), 均表现为弱致病力的菌株有5个(DI<20);不同芝麻品种对不同菌株的抗性有一定差异。该方法可应用于芝麻枯萎病病原菌致病性测定和芝麻种质抗枯萎病特性评价, 并为后续的机理研究提供了技术支持。