转抗菌肽基因毛白杨的培育及其对溃疡病的抗性

以毛白杨为转基因受体材料,利用根癌农杆菌介导法转化来源于益母草的阳离子抗菌肽基因LJAMP2。经卡那霉素筛选,共获得50株抗性植株。GUS组织染色和PCR检测显示有30株抗性植株呈阳性,初步证明外源目的基因已整合到毛白杨基因组中。RT-PCR证实抗菌肽基因LJAMP2在转基因植株中能大量表达。离体抗病性试验表明:转基因毛白杨细胞粗提液的抑菌能力明显强于非转基因植株。进一步将溃疡病菌接种在转基因和野生型毛白杨茎段上培养30天,转基因植株的病级指数均低于非转化植株。上述抗性试验结果表明:在毛白杨中超量表达益母草抗菌肽基因LJAMP2能显著提高其溃疡病抗病性。     

Fused: a promising molecular target for an RNAi-based strategy to manage Bt resistance in Plutella xylostella (L.)

Bacillus thuringiensis (Bt), a naturally occurring entomopathogenic soil bacterium, has been the active ingredient of sprayable Bt biopesticides for over a century. Insecticidal Bt Cry proteins are particularly well suited for use as plant-incorporated protectants in transgenic crops (Bt crops) due to their specificity against insect pests and safety for non-target organisms. The sustainability of these Bt products, however, has been challenged by the development of resistance in the field. RNA interference (RNAi), a species-specific control alternative that has been deregulated and commercialized in the USA and Canada, provides a new mode of action to complement the existing Bt products. Based on our preliminary research, we hypothesized that pyramiding/integrating Bt with RNAi can address/manage resistance issues related to Bt traits. To examine this overarching hypothesis, we (1) cloned and characterized a serine/threonine kinase gene (fused) of the Hedgehog (Hh) signaling pathway in the diamondback moth, Plutella xylostella (L.), a global superpest; (2) profiled Pxfused expression in Bt-resistant P. xylostella strains; and (3) investigated the involvement of Pxfused in Bt Cry1Ac resistance in P. xylostella. Pxfused expression was elevated ubiquitously in all Bt-resistant strains, and silencing of Pxfused led to larval and pupal mortality in both Cry1Ac-susceptible and -resistant strains, suggesting that Pxfused is a potential target for RNAi-based resistance management. Taken together, our results not only identify a molecular target to control a devastating lepidopteran pest, but also shed light on a novel resistance management strategy through the integration of two biotechnological techniques with distinct modes of action.

     

杨树转基因研究进展及展望

杨树是重要的栽培树种,也是研究林木基因工程的重要模式植物。杨树转基因研究可以打破种属限制,具有高效性和专一性的特点,是对杨树进行遗传改良的重要手段。为了更好的进行该方面的工作,本文介绍了杨树转基因涉及到的抗虫、抗除草剂、木材材性改良、抗逆、抗病、激素调控、开花调控和植物修复等应用领域的研究进展及现状,分析了影响杨树农杆菌转化效率的主要因素,并探讨了杨树转基因研究存在的问题及发展方向,希望能为后期从事林木基因工程研究的科研工作者提供依据。     

杨树抗性转基因研究进展

本文重点介绍了杨树基因工程中采用的抗虫基因、抗病基因、抗除草剂基因和抗逆境基因类型及研究现状 ,并对杨树抗性转基因研究存在的问题及解决途径作了探讨     

杨树基因工程的研究进展

围绕转基因研究着重论述了杨树在抗病虫害育种、抗除草剂、抗非生物逆境胁迫、木质素结构调整以及转基因树木安全性等方面的研究进展,并对转基因杨树的应用前景作了展望。     

转基因林木研究进展

着重论述了林木抗虫,抗菌,抗除草剂,生殖发育调控,抗逆境,材性改良等转基因林木的研究进展,简述了转基因林木的应用现状,提出了林木基因工程现存的问题,并对该领域进行了展望。     

意大利的杨树病害及其防治

本文简要介绍了意大利杨树病害的发展历史、现状、几种主要的病害以及围绕病虫害防治所做的一系列抗性育种工作。本文旨在杨树抗病害方面学习意大利的先进经验,以他人走过的弯路作为借鉴。     

植物基因工程及其在林木害虫防治上的应用

植物基因工程主要包括目的基因的分离、基因工程载体的构建、植物细胞的遗传转化、转化细胞的组织培养和植株再生、外源基因表达的检测等几个方面。利用该技术目前已转化了20多种林木树种,并成功地将抗虫的苏云金芽杆菌内毒素基因和蛋白酶抑制剂基因转入林木树种。     

TERF1基因增强糖橙抗病性的初步研究

将获得转TERF1基因糖橙的春梢叶片进行柑桔溃疡病和炭疽病的接种.初步试验表明,TERF1基因增强了糖橙的广谱抗病性.在接种溃疡病后,转基因植株的平均发病率下降了50%左右,且发病的时间比冰糖橙和对照推迟了7 d,其中3号株系的抗病性最强,叶片无病症.柑橘叶片接种炭疽病后,转基因植株的叶片与对照相比基本上表现出针孔处病斑减小,延迟发病等现象.其中3号和11号转TERF1基因糖橙叶片上根本观察不到炭疽病的典型病斑,且叶片也无褪绿.     

抗虫转基因欧洲黑杨的Western印迹法分析

抗虫转基因欧洲黑杨的Ｗｅｓｔｅｒｎ印迹法分析陈颖，李强，李玲，韩一凡，田颖川（中国林业科学研究院林业研究所北京１０００９１）（中国科学院微生物研究所北京１０００８０）关键词Ｂｔ．基因，转基因欧洲黑杨，Ｗｅｓｔｅｒｎ印迹法ＷＥＳＴＥＲＮＢＬＯＴＡＮＡＬ...     

过量表达LAR3和Bbchit1以提高毛白杨对叶枯病的抗性

利用转基因技术将多种抗病基因共同转入毛白杨中以提高其抗性,从而获得毛白杨抗病新品种是目前解决杨树真菌病害的主要研究方向之一。本研究通过根癌农杆菌介导的二次遗传转化,将来源于球孢白僵菌几丁质酶基因Bbchit1转入过量表达无色花色素还原酶基因LAR3转基因毛白杨中,实时定量PCR显示Bbchit1与LAR3均能有效表达,离体抗病试验显示Bbchit1+LAR3共表达转基因毛白杨细胞粗提液对杨树叶枯病菌具有明显抑制作用,进一步将叶枯病菌接种在转基因和非转基因毛白杨叶片上培养30天,转基因植株的感病面积均低于非转基因植株且Bbchit1+LAR3共表达转基因株系抗病效果更明显。上述抗病试验结果表明:LAR3和Bbchit1在杨树中共表达可提高其对叶枯病的抗性。     

近10年我国林木抗虫性研究概述

过去10年里,为了有效地解决我国人工林大范围严重的害虫危害,人们对林木的抗虫性问题,从概念、分类分级方法,到林木抗虫机制、抗虫力与耐害力、个体抗性与群体（林分）抗性等各个方面,进行了大量的研究,取得了许多成果。文中对此进行了分类整理,同时对一些相关概念做了初步的界定。     

Electrical Capacitance and Resistance Assessing Tree Vigor

When individual trees or large areas of forests are subjected to environmental stresses, such as drought, cold, repeated insect pests and diseases, decline of tree vigor and forests degradation may take place. For forest managements, it is necessary to predict the health conditions of individual trees or forests in both outbreak and nonoutbreak areas. Thus, forest resources can be better protected and risks of environmental stresses can be reduced. Meanwhile, in the field of forest theory rese…     

白桦转基因遗传改良研究进展

随着科学技术深入发展, 转基因技术已经成为林木定向遗传改良的重要途径之一, 林木基因工程也随着转基因技术的成熟而步入快速发展时期。目前国内对白桦(Betula platyphylla)的研究处于常规育种向分子育种的过渡阶段, 转基因技术在白桦定向培育过程中的应用才刚刚起步。文中从白桦开花、抗虫、木材改良、抗非生物胁迫等方面总结了国内外白桦转基因研究现状, 对转基因技术在白桦遗传改良种应用中存在的问题进行探讨, 展望了转基因技术在白桦遗传改良中的应用前景, 以期为白桦基因工程研究提供参考。     

Defensin (TvD1) from Tephrosia villosa exhibited strong anti-insect and anti-fungal activities in transgenic tobacco plants

Plant defensin is a small, cationic, cysteine-rich broad-spectrum antimicrobial peptide with four or five disulfide bridges and has been shown to be a component of the innate immunity system in plants. In the present study, the defensin gene (TvD1) from Tephrosia villosa was overexpressed in tobacco and characterized. Two high-expression (T1, T26) and one low-expression (T13) plant lines were selected through semi-quantitative RT-PCR and used for bioassays along with non-transgenic controls. The high-expression plant line exhibited strong in vivo anti-fungal and anti-feedant activity against the pathogen Rhizoctonia solani and the first and second instar larvae of the Spodoptera litura (F.), the tobacco cutworm, respectively. The low-expression plant line showed a moderate level of tolerance/resistance in the bioassays. The recombinant peptide (rTvD1) exhibited toxicity to tobacco pollen grains in the germination assay, but transgenic plants produced copious fertile pollen and set capsules with viable seeds. The results of this study demonstrate that the single gene (TvD1) effectively controls both fungal and insect pests and, hence, it can be used for crop transformation.     

表达豇豆胰蛋白酶抑制剂转基因杨树的虫试试验

将转豇豆胰蛋白酶抑制剂基因的三倍体毛白杨回交杂种 [(毛新杨×毛白杨 )×毛白杨 ]]于春季萌发前截干 .用萌条上的树叶对 3种主要杨树害虫 :天幕毛虫 (Malacosomadisstria)、舞毒蛾 (Lymant riadisparL .)和杨雪毒蛾 (StilpnotiacandidaStaudinger)进行离体虫试试验 ,年底测量各转基因株系萌条树高和地径 ,以衡量其生长性状 .结果发现 ,与对照相比 ,转基因株系的树高和地径没有显著降低 ,说明外源基因的导入没有影响转基因杨树的速生特性 ,相反 ,TG0 4的树高和地径较对照有明显的提高 ,这可能是外源导入后促进转基因杨树生长的结果 ;转基因植株均能显著提高害虫的死亡率 ,降低幼虫的排粪量、蛹重、体重增加量和成虫的产卵量 ;其中 ,TG0 7、TG0 8和TG713个转基因株系的抗虫特性较为突出 ,表明CpTI基因在它们的树体内表达较为活跃和稳定     

Towards a resistance management strategy for Helicoverpa armigera in Bt-cotton in northwestern China: an assessment of potential refuge crops

Transgenic Bt-cotton now dominates the cotton-growing belt in northwestern China where there are few natural plant refuges to act as sources of moths susceptible to Bt toxin. As an initial step towards developing an insect resistance management (IRM) plan for the pest moth, Helicoverpa armigera, on Bt-cotton in this part of China, we assessed the potential of six crops grown in two configurations to act as refuge hosts for susceptible moth production in both broad-acre and small-holder farms. Egg and larval abundance indicated that H. armigera had a preference for chickpea, pigeon pea, and corn, over cotton, sorghum, and benne (sesame). There were no significant differences in egg or larval abundance between plot and strip configurations of these crop hosts. We found that sorghum was not attractive to H. armigera, contrary to the findings in other cotton production areas. Moreover, chickpea, pigeon pea, and corn were determined to be potential refuge hosts based on egg and larval abundance over the growing season. From the adult moth population dynamics in broad-acre and small-holder farms, the efficiency of alternative refuge crops, and local agricultural practices, we recommend that spring corn be grown as a strip crop as part of an IRM strategy to improve the probability of Bt-susceptible moth production and mating with resistant moths in broad-acre farms. In small-holder farms where current agricultural practices are entrenched, wheat and summer corn should be evaluated as refuge crops in the future.