转基因小麦沉默的HMW-GS基因的遗传及表达

为了研究转基因小麦QQ5沉默的HMW-GS基因的遗传规律.以QQ5与育成品种高原314和新春13号进行正反交,再用育成品种进行曰交,采用SDS-PAGE技术检测并分析各组合亲本、F1、F2、BC.F1的HMW-GS组成.结果表明,HMW-GS基因沉默表现为显性,且能稳定遗传,杂交及回交后代符合3:1和1 : 1的分离比,遵循孟德尔遗传方式.     

转基因大麦的gfp基因表达及染色体定位

为了深入探讨转基因的整合位置对基因表达的影响，对转绿色荧光蛋白基因（gfp）大麦的荧光表达和gfp基因的染色体位置进行了研究。结果表明，绿色荧光蛋白（gfp）在大麦的根尖和花粉中均有表达，四倍体的荧光表达强度大于二倍体，表现出基因的剂量效应。gfp基因插入到第6染色体（6H）短臂和另一染色体的短臂近末端。     

转基因小麦外源基因在主栽小麦种质中的遗传规律

为了研究转基因小麦中外源品质基因1D x 5在我国主栽小麦种质中的遗传规律,以转基因小麦B 72-8-11b为父本,主栽品种川89-107和鄂麦18为母本进行杂交,采用SDS-PAGE技术检测并分析各组合亲本、F1、F2、BC1F2、BC2F1、BC2F2代的HMW-GS组成。结果表明,外源基因有效地整合到主栽小麦的基因组中,并能够遵循孟德尔遗传模式稳定地遗传给下一代。     

大豆炸荚基因GmAGL8的转基因鉴定与功能初探

炸荚是大豆的一种自然特征属性,是影响大豆产量的重要因素之一。本研究以前期获得的转大豆炸荚相关基因GmAGL8的T_1代植株为材料,继续繁育获得T_2和T_3代;采用PCR和RT-q PCR检测方法对转基因植株进行基因遗传稳定性和表达情况分析;并以野生型中黄10号为对照,对大豆炸荚性状进行了鉴定分析。结果表明:转基因植株阳性率T_1代为87.5%,T_2和T_3代均达到100%,说明GmAGL8基因已基本能够在转基因后代中稳定遗传。RT-q PCR检测结果显示,转基因植株中GmAGL8基因的相对表达量都明显高于非转基因植株,且各转基因植株之间表达量具有差异性。对T_1、T_2和T_3代植株炸荚率进行了统计,不同世代转基因大豆的平均炸荚率为9.09%,而非转基因大豆炸荚率为83.3%,转基因与非转基因大豆之间炸荚率存在显著差异。综上所述,GmAGL8基因已基本实现在大豆转基因后代中稳定遗传并正常表达,表型鉴定结果初步证明了GmAGL8基因与大豆炸荚性状相关。     

抗除草剂转基因水稻稳定系TR4的获得及其遗传研究

 用Bar基因通过基因枪法轰击京引119未成熟胚,得到抗除草剂Basta的转基因植株,经加代和选择得到抗除草剂的稳定系TR4,其基本农艺性状与京引119相仿。用TR4与4个粳稻品种杂交进行遗传分析表明,TR4的抗除草剂特性由一对显性基因控制,符合孟德尔遗传规律。该外源基因可通过有性杂交转育,在杂交稻和常规直播稻的选育上有较大的利用价值。     

GFP绿色荧光法辅助筛选玉米转基因植株

利用GFP(Green Fluorescence Protein)绿色荧光蛋白基因作为辅助性筛选标记基因,基因枪法轰击玉米自交系A188和Qi319未成熟胚的胚性愈伤组织。瞬间表达实验结果表明,轰击后GFP表达可持续近20 d,轰击后10 d左右表达量仍保持在较高的水平。采用潮霉素筛选、GFP荧光法鉴定,获得一批玉米转基因植株,转化频率为0.33%~0.47%。分子鉴定和后代分析的结果表明,外源基因已经整合到了玉米基因组中,并能够稳定地表达和遗传。GFP作为辅助性筛选标记基因可以直观、快速和有效地消除玉米遗传转化工作中的假阳性现象。     

一种可视化快速筛选转基因玉米的方法研究

利用可视化表型分析快速筛选转基因后代可以有效提高遗传转化和基因功能的验证效率.本文研究一种高效、低成本的筛选转基因玉米的新方法,该方法利用愈伤组织和糊粉层特异性表达启动子Ltp2和可视化红色荧光蛋白编码基因DsRed构建植物表达载体pCAMBIA3300-DsRed,通过农杆菌介导法侵染玉米HiⅡ幼胚,诱导出分子检测为...     

温室条件下抗除草剂转基因水稻与杂草稻杂交和回交后代的适合度分析

在温室条件下,将抗除草剂转基因水稻Y0003和99-1与马来西亚杂草稻和国内安徽塘稻人工授粉获得的携带抗性基因的F1、F2与相应杂草稻回交,统计回交结实率和携带抗性基因的比例,并对携带抗性基因的杂交/回交代的适合度进行了测定,综合判断抗性基因漂移的风险。研究表明,在人工授粉的条件下,携带抗性基因的杂交代都能与相应的杂草稻回交并结实,结实率为15%～60%。F1的种子绝大多数都对草丁膦表现出了良好的抗性,F2和回交代分别表现为3∶1和1∶1的抗性分离比例,符合孟德尔遗传规律。适合度研究表明,杂交代、回交代与相应的杂草稻相比没有明显差异,大多数的杂交种的适合度和回交代相差不大,个别杂交代的适合度没有回交代高。以上研究结果表明在无除草剂选择压下,携带转基因水稻Y0003和99-1的抗性基因的2种杂草稻的杂交和回交后代有在适宜环境条件下生存定植的可能性。     

几个转bar基因水稻外源基因的遗传学行为研究

利用春江03等4个品种的转基因株及后代稳定系进行遗传研究表明,bar基因作为一个显性单位已整合在水稻核基因组中,转基因株稳定系TR3、TR4、TR5、TR6对除草剂Basta的抗性均受1对显性基因控制,且4个转基因株稳定系间bar基因互不等位,不同位点的bar基因之间存在基因互作,表现为基因重叠作用。Southern杂交分析结果表明,TR5、TR6的bar基因整合位点互不相同,但可通过杂交育种重组到同一个水稻核基因组中。     

小麦隐性核不育保持系表达载体的构建及拟南芥遗传转化

利用小麦杂种优势能够创制稳定的雄性不育系。相比于细胞质雄性不育系和光温敏雄性不育系,细胞核雄性不育具有育性稳定、易恢复、不受环境影响等特点。小麦隐性核不育基因 ms1的克隆以及杂交制种技术（hybrid seed production technology,SPT）的开发,为雄性不育的保持和杂种优势利用奠定了基础。本研究将小麦花粉育性恢复基因 Ms1、花粉致死基因 ZmAA1、红色荧光蛋白基因 DsRed2及其特异性启动子和终止子连接到表达载体pGEII上,转化野生型拟南芥,得到6株转基因拟南芥。通过荧光显微镜观察拟南芥种子,发现野生型拟南芥种子表面平滑,不能发出红色荧光;而转基因种子表面呈现颗粒状,能够发出红色荧光。这说明表达载体构建成功,并能够在拟南芥中成功表达。这为创制人工小麦隐性核不育系奠定了理论和材料基础。     

Host suitability of three avocado cultivars (Persea americana Miller: Lauraceae) to oriental fruit fly (Bactrocera (invadens) dorsalis (Hendel) (Diptera: Tephritidae))

The oriental (invasive) fruit fly Bactrocera (invadens) dorsalis (Hendel) (Diptera: Tephritidae) has invaded Africa and is currently established in over 30 countries and has caused in massive fruit losses. Avocado (Persea americana Miller) is among the fruit with which the pest is associated. The aim of the study was to determine the host status of a ‘black-skinned’ cultivar “Hass” and two ‘green-skinned’ cultivars “Pinkerton” and “Fuerte” to B. (invadens) dorsalis. In the laboratory study conducted in Kenya in 2012 and 2013, punctured and unpunctured recently-harvested fruit were exposed to sexually-mature fruit fly in ‘no-choice’ experiments. B. (invadens) dorsalis readily developed in the punctured fruit but not in the uncompromised fruit (fruit not damaged by insects or handling). This result was confirmed in Tanzania in 2013 where flies were introduced to unpunctured fruit. In the field studies the fruit of each cultivar were caged in situ with flies for 72 h. On dissection only fruit damaged by false codling moth (Thaumatotibia leucotreta (Meyrick)) were found to harbor B. (invadens) dorsalis. In the light of these results the risk imposed for “Hass”, “Pinkerton” and “Fuerte” by B. (invadens) dorsalis is negligible under standard export conditions. The findings of this study and the implications to regional and international trade are discussed.     

Solvent transport phenomenon of cellulose microfiber (CMF) reinforced poly (ethylene-co-vinyl acetate) (EVA) composites

Composite films based on EVA with CMF were prepared and characterized for their solvent transport nature. CMF were derived from Hibiscus sabdariffa by steam explosion technique and incorporated in the EVA matrix by extrusion. Solvent transport by the composites was analyzed using hexane, heptane and octane as penetrant molecules. The solvent uptake behavior of EVA/CMF composites is explained on the basis of the penetrant size, temperature and the filler loading. The composite had improved barrier property compared with unfilled EVA. This in turn is an indication of good fiber-matrix interaction in the composite system. Diffusion parameters such as diffusion coefficient, permeation coefficient and Arrhenius activation parameters and thermodynamic parameters such as enthalpy and entropy were evaluated in detail. The mode of transport was found to be Fickian.     

Thermal degradation behaviors and fire retardant properties of poly(1,3,4-oxadiazole)s (POD) and poly(m-phenylene isophthalamide) (PMIA) fibers

Thermal degradation behaviors and fire retardant properties of poly(1,3,4-oxadiazole)s (POD) and poly(m-phenylene isophthalamide) (PMIA) fibers were investigated. The thermal gravimetric analysis (TGA) demonstrated that POD exhibited higher onset thermal degradation temperature (T_onset) than PMIA, exceeding nearly 80 °C. The thermal degradation kinetics, evaluated by the modified Coats-Redfern method, displayed that the apparent activation energy (E_a) of POD and PMIA fibers was similar when the conversion rate (α) ranges from 0.2 to 0.5, while with the α from 0.6 to 0.8, the E_a of POD was significantly lower than that of PMIA. The fire retardant performance of POD and PMIA fibers were evaluated by cone calorimeter under heat fluxes of 35, 50 and 75 kW/m², during which the temperature of the fibers were monitored by a thermocouple. Surprisingly, POD fibers showed inferior fire retardant performance in comparison with PMIA, with lower time to ignition (TTI) and higher peak heat release rate (PHRR). The origin of the different fire retardant properties of both fibers was revealed by analyzing the residual chars and gaseous products during thermal pyrolysis. The morphology confirmed that stable and compact chars can be formed in PMIA. In addition, the Fourier Transform Infrared Spectroscopy (FTIR) characterization of the residual char revealed that POD can form carbonaceous chars at the heat flux of 50 kW/m², while the heat flux of PMIA was 75 kW/m². The pyrolysis products characterized by pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS) indicated that POD can be pyrolyzed completely at 600 °C, while the temperature of PMIA was 700 °C.     

Preparation of poly(vinyl acetate)/clay and poly(vinyl acetate)/poly(vinyl alcohol)/clay microspheres

Poly(vinyl acetate) (PVAc)/poly(vinyl alcohol)(PVA)/montmorillonite (MMT) clay nanocomposite microspheres with a core/shell structure have been developed via a suspension polymerization approach. In order to prepare the PVAc/MMT and PVAc/PVA/MMT nanocomposite microspheres, which are promising precursor of PVA/MMT nanocomposite microspheres, suspension polymerization of vinyl acetate with organophilic MMT and heterogeneous saponification were conducted. A quaternary ammonium salt, cetyltrimethylammonium bromide, was mixed with the MMT in the monomer phase prior to the suspension polymerization. The rate of conversion decreased with an increase in MMT concentration. The incorporation of MMT into the PVAc was verified by FT-IR spectroscopy. Organic vinyl acetate monomers were intercalated into the interlayer regions of organophilic clay hosts and followed by suspension polymerization. Partially saponified PVA/MMT nanocomposite microspheres with a core/shell structure were successfully prepared by heterogeneous saponification.     

Suppression of green peach aphid (Myzus persicae) (Sulz.), citrus mealybug (Planococcus citri) (Risso), and two spotted spider mite (Tetranychus urticae) (Koch.) attacks on tomatoes and cucumbers by aqueous extracts from vermicomposts

Vermicomposts are produced through interactions between earthworms and microorganisms in the breakdown of organic wastes. Aqueous extracts were prepared in commercial brewing equipment (Growing Solutions Inc.) from vermicomposts processed from pre-consumer food waste. The ratio of vermicompost to water was 1 to 5 v:v to produce a 20% aqueous solution. The effects of soil drenches at dilutions of 20%, 10%, and 5% vermicompost extracts, were compared with those of deionized water, in the suppression of green peach aphids, mealybugs, and two spotted spider mites attacking tomatoes and cucumbers, in greenhouse cage experiments.     

Nemato-toxic potential of Betel (Piper betle L.) (Piperaceae) leaf

In vitro and in-planta trials were conducted to determine the nemato-toxic potential of aqueous leaf extracts of Betel (Piper betle L.) against the root-knot nematode species, Meloidogyne incognita, at five concentrations, standard extract (S) [1:2 (w/v] and its four dilutions, 20%, 40%, 60%. 80% of S. In the in vitro trials, second-stage juveniles (J₂s) and eggs of M. incognita were directly exposed to the extract while in the in-planta trials, the effect of the extract was evaluated as a root dip treatment using tomato plants. The findings indicated that the extract was lethal to J₂s but also inhibited egg hatch. The seedling dip treatments reduced root infestations in terms of gall formation, egg production and J₂ population densities in soil while simultaneously enhancing the growth of tomato plants. All these effects varied in a dose-dependent manner. Based on the LC₅₀ value, the eggs were found to be less sensitive to the extract than J₂s. One hundred percent of mortality of J₂s was recorded at four concentration levels, 40%, 60% and 80% of S and S, while 100% egg inhibition was only recorded at S. The highest reductions in gall formation (i.e., number of galls/root system), egg production and J₂ population were all recorded at S as 83%, 87% and 84% compared to the untreated water control. The maximum growth enhancement, which was 235% of the control, was detected in the root length of treated tomato plants. It appears the aqueous leaf extract of P. betle shows promise as a sustainable eco-friendly nematicide for the management of M. incognita.     

Biological study of Plutella xylostella (L.) (Lep: Plutellidae) and it's solitary endoparasitoid, Cotesia vestalis (Haliday) (Hym. Braconidae) under laboratory conditions

Plutella xylostella (L.) (Lep: Plutellidae), is a destructive pest of brassicaceous crops in the world. Cotesia vestalis (Haliday) is one of most important biological control agents of P. xylostella in the world and Iran. Both of P. xylostella and C. vestalis biology were carried out in laboratory condition. Results showed that development time of immature stages of P. xylostella including egg, Instar I, Instar II, Instar III, Instar IV, prepupa, pupa were 2.39 +/- 0.17, 2.18 +/- 0.17, 2.06 +/- 0.28, 2.14 +/- 0.14, 2.54 +/- 0.12, 0.40 +/- 0.12 and 4.23 +/- 0.23 days, respectively. Longevity of female and male were 28.26 +/- 0.05 and 30.22 +/- 0.05 days. By dissecting the parasitized larvae, the egg incubation period of C. vestalis was recorded 1.73 +/- 0.06 days. In long-term oviposition trials, females laid eggs on P. xylostella larvae for up to 10 days. Larval development of the parasitoid in host only required 6.47 days: the first instar larva required 3.25 +/- 0.047 days; the second instar larva needed 2.78 +/- 0.1 days and the third instar larvae exited the host and pupated in, 0.4 +/- 0.07 days. Prepupal and pupal period of wasp were 1.9 +/- 0.06 and 2.13 +/- 0.09 day, respectively. Unmated female and male longevity of wasp were 16.83 +/- 0.37, 16.25 +/- 0.17 and sex ratio is male-biased. When a mixed group and isolated of instars were presented for parasitoid, the 2nd and 3rd instar larvae were so preferred and the 4th instar was less attractive for selection. In choice experiment, the percentage parasitism of 2nd, 3rd and 4th instars was 78.58, 69.94 and 4.36%, respectively. The rapid oviposition rate, short life duration and high percentage parasitism increases parasitoid potential for suppression of host population. Present results suggest that C. vestalis has considerable potential as a biological control agent for P. xylostella.     

Can spinosad-resistant Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) be managed with spinosad and predatory mites (Acari)?

Frankliniella occidentalis (Pergande) is a serious pest of a wide range of horticultural and ornamental crops. Populations resistant to most conventional insecticides, including–spinosad, have been detected. To control spinosad-resistant thrips, growers could use a ‘high-rate’/biological control strategy. The proposed strategy is based on a single application of spinosad at double the recommended application rate followed by releasing predatory mites (Acari), which are used as biological control agents of F. occidentalis. This study compared two resistance management strategies on a spinosad-resistant F. occidentalis strain: applying spinosad at twice the recommended rate, and spraying at twice the rate then releasing predatory mites, Typhlodromips montdorensis (Schicha), Neoseiulus cucumeris (Oudemans) and Hypoaspis miles (Berlese). Direct exposure to twice the recommended rate of spinosad killed 100% of all adults of all species of predatory mites. Spinosad residues aged 2–48 h were also highly toxic to adults of all three mite species, causing 96–100% mortality. Spinosad residues aged 48–168 h were less toxic to N. cucumeris than to T. montdorensis and H. miles. LT₂₅ of double the recommended rate of spinosad for T. montdorensis, N. cucumeris and H. miles were calculated as 6.02, 5.3, and 7.08 days, respectively. When released after applying spinosad, T. montdorensis was the most successful species in reducing thrips numbers, followed by N. cucumeris and H. miles. By releasing mites 6–7 days after a spinosad application, our results suggest that F. occidentalis can be effectively controlled. The practical implications of implementing a ’high-dose/biological control’ strategy are discussed.