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国家高技术研究发展计划(2008AA10Z414)

作品数:8 被引量:59H指数:3
相关作者:周明国王建新徐建强张聪章四平更多>>
相关机构:南京农业大学河南科技大学更多>>
发文基金:国家高技术研究发展计划国家重点基础研究发展计划国家自然科学基金更多>>
相关领域:农业科学化学工程更多>>

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8 条 记 录,以下是 1-9
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一种经济高效的禾谷镰孢菌原生质体转化方法
禾谷镰孢菌(Fusarium graminearum)原生质体遗传转化技术是建立禾谷镰孢菌突变体文库及研究其功能基因组的一种重要方法,本文以禾谷镰孢菌Y-A为供试菌株,研究酶解材料、酶组合、酶浓度、酶解时间、酶解温度和最...
侯毅平陈长军王建新周明国
关键词:禾谷镰孢菌原生质体制备
禾谷镰孢菌微管相关蛋白基因(map)克隆及其与多菌灵抗药性关系分析被引量:1
2009年
根据禾谷镰孢菌(Fusarium graminearum)测序菌株NRRL31084(PH-1)的微管相关蛋白基因(map)核苷酸序列设计4对引物,采用PCR方法测定了禾谷镰孢菌对多菌灵(MBC)不同敏感性表型的6个中国菌株的map全序列。DNA序列比对表明,中国的3个敏感菌株和3个抗药菌株的map核苷酸序列相似性没有差异,表明多菌灵抗药性与微管相关蛋白无关。该基因全长1 793 bp,含有6个内含子,编码199个氨基酸;与NRRL31084的map核苷酸序列相似性为99.39%,存在10个差异核苷酸,与其所编码的氨基酸序列相似性为100%。
陈长军李俊于俊杰王建新周明国
关键词:禾谷镰孢菌多菌灵抗药性
Biological Mode of Action of Dimethomorph on Pseudoperonospora cubensis and Its Systemic Activity in Cucumber被引量:6
2009年
Dimethomorph is a fungicide with high activity against Peronosporomycetes plant pathogens. The present study showed that dimethomorph is effective on controlling the oomycete fungal pathogen Pseudoperonospora cubensis causing downy mildew on cucumber. The fungicide did not affect zoospores discharge from sporangia of P. cubensis, but it strongly inhibited mycelial growth and sporangial production in vitro and increased lysis of zoospores. Dose of 2 mg L^-1 of dimethomorph was sufficient to inhibit mycelial growth and sporangial production of P. cubensis on leaf disks, 5 mg L^-1 was enough to lyse zoospores of P. cubensis, and 25 mg L^-1 was required to inhibit sporangial production on detached leaves. In whole plant tests, dimethomorph exhibited strong protective and curative activity. Dimethomorph when applied at a dose of 300 mg L^-1 for 1, 3, 5, 7 days before inoculation exhibited 100% efficacy on disease control. On the other hand, efficacies of 67.1 and 31.5% were obtained when the same dose of dimethomorph was applied for 1 and 3 days after inoculation, respectively. So dimethomorph had persistence effect on leaves for 7 days at least and exhibited strong protective and curative activity. Bioassay analyses showed that dimethomorph could be translocated in the xylem system, redistributed in the leaf, and penetrated from the upper surface to the lower surface of the leaf but could not be translocated in phloem system or transferred from the roots to leaves of cucumber plants in sufficient amounts for disease control. The biocharacteristics of dimethomorph make it well suitable for integration of a control programme against downy mildew disease on cucumber and as a component to delay other peronosporomycetes fungicide-resistance development.
WANG Han-cheng ZHOU Ming-guo WANG Jian-xin CHEN Chang-jun LI Hong-xia SUN Hai-yan
关键词:DIMETHOMORPH
玉蜀黍赤霉有性重组体的诱导及其生物学特性
2009年
本研究以硝酸盐营养缺陷型突变体(nit)和多菌灵抗性为遗传标记,在5个所选菌株中设计了3个玉蜀黍赤霉病菌Gibberella zeae的杂交组合,使各菌株之间进行杂交,从而诱导有性重组体。从各杂交组合的后代中任意挑选出3个有性重组体,比较了这些有性重组体与其亲本在无性和有性阶段的主要生物学性状。结果表明,玉蜀黍赤霉中的nit基因及对杀菌剂多菌灵的抗药性基因可以通过有性杂交的方式重组,即发生了有性重组。有性重组体与其亲本在菌落生长、培养性状和致病性方面没有显著差异;但某些有性重组体中,产孢量和产子囊壳能力方面存在一定差异。总体看来,有性重组体仍然保持了较高的适合度。因此,可以认为有性重组在玉蜀黍赤霉群体对多菌灵抗药性发生发展以及群体遗传进化中可能起着重要的作用。
陈雨黄婷婷周明国
关键词:NIT突变体生物学特性遗传进化
Activity of Azoxystrobin and SHAM to Four Phytopathogens被引量:2
2009年
The study was conducted to make clear the activity of azoxystrobin to 4 plant pathogens and the synergistic effects of salicylhydroxamic acid (SHAM), which acted on the alternative oxidase. It was also conducted to be aware of the mechanism of azoxystrobin in inhibition on mycelial respiration and the influence of SHAM. The activity test of azoxystrobin and SHAM was carried out with a mycelial linear growth test and spore germination test. Other related biological properties were also observed. Inhibition of azoxystrobin and SHAM on 4 pathogens was determined by using SP-II oxygraph system. Azoxystrobin inhibited mycelial growth in Colletotrichum capsici, Botrytis cinerea, Rhizoctonia solani, and Magnaporthe grisea, respectively; it also inhibited conidia germination, and conidia production in C. capsici, B. cinerea M. grisea, and sclerotia formation in R. solani. Moreover, it created stayed pigment biosynthesis in C. capsici and M. grisea somehow. Salicylhydroxamic acid enhanced inhibition by azoxystrobin. An oxygen consuming test of the mycelia showed that azoxystrobin inhibited all the 4 fungi's respiration in the early stages. With the concentration rising up, the effectiveness increased. However, as time went on, the respiration of the mycelia treated with fungicides recovered and SHAM could not inhibit the oxygen consuming. This reaction between the mycelia and the fungicides appeared not to initiate alternative respiration but rather the other mechanism created a lack of efficacy.
JIN Li-hua CHEN Yu CHEN Chang-jun WANG Jian-xin ZHOU Ming-guo
枯草芽孢杆菌生防菌株NJ-18的发酵条件优化被引量:42
2010年
通过单因子试验筛选到了适合枯草芽孢杆菌NJ-18生防菌株生长及其拮抗物质产生的最佳碳源为玉米粉和麦芽糖,最佳氮源为豆粕,最佳金属离子是Ca2+。用正交试验法确定了该培养基各组分的最佳含量为:玉米粉10g.L-1、麦芽糖30g.L-1、豆粕30g.L-1、CaCO31.5g.L-1;在确定适合该菌株的培养基组分基础上,对其培养条件进行优化试验,结果表明:初始pH为6.8、培养温度为28℃、250mL摇瓶装液量为90mL、接种量为2%(体积分数)、培养时间为36h、转速为180r.min-1时为该菌株的最佳培养条件。
章四平刘圣明王建新周明国
关键词:单因子试验枯草芽孢杆菌发酵条件拮抗物质
禾谷镰孢菌β_2-微管蛋白在大肠杆菌中的可溶性表达及纯化被引量:4
2012年
【目的】在大肠杆菌中表达禾谷镰孢菌(Fusarium graminearum)的β2-微管蛋白,筛选使β2-微管蛋白可溶性表达的诱导因子,并对可溶性蛋白进行纯化。【方法】以构建好的质粒(pET32a+-β2-tubulin)为模板,PCR扩增出β2-微管蛋白基因,将其克隆到pET30a+表达载体上,并转化到表达宿主菌BL21(DE3)及Rossatta(DE3)pLysS,筛选阳性克隆,进行蛋白诱导表达,并筛选蛋白可溶性表达的诱导因子;对纯化后的蛋白进行SDS-PAGE和Western blot验证。【结果】获得了在大肠杆菌中表达效率高的阳性克隆;为了克服常规诱导条件下表达的β2-微管蛋白主要以包涵体形式存在的问题,通过对诱导温度、时间、诱导物浓度、初始诱导时的菌液浓度、培养液及宿主菌等六因子的综合分析,获得了β2-微管蛋白可溶性表达的优化条件;利用HisTrap HP预装柱对β2-微管蛋白进行纯化,可获得纯度很高的可溶性β2-微管蛋白;SDS-PAGE确认表达出的融合蛋白分子量为51.6 kD;Western blot证实表达的融合蛋白能与6×His及β-微管蛋白的单克隆抗体发生特异性反应。【结论】本研究为外源基因在大肠杆菌中的可溶性表达提供了参考;获得的微管蛋白为研究微管蛋白靶标类药物的抗性机制以及生产中开发新的以微管蛋白为靶标的杀菌剂提供了物质基础。
徐建强周裕军张聪周明国
关键词:禾谷镰孢菌包涵体可溶性表达纯化
Effect of Azoxystrobin on Oxygen Consumption and cyt b Gene Expression of Colletotrichum capsici from Chilli Fruits被引量:2
2009年
Azoxystrobin acts as an inhibitor of electron transport by binding to the Qo center of cytochrome b (cyt b). Resistance to azoxystrobin was usually caused by the point mutation of cyt b gene or by the induction of alternative respiration. Oxygen consumption test for mycelia of Colletotrichum capsici showed that azoxystrobin inhibited mycelial respiration within 12 h; however, as time went on, the respiration of the mycelia recovered when the mycelia were treated with azoxystrobin and salicylhydroxamic acid (SHAM, a known inhibitor of alternative respiration), and the oxygen consumption of the mycelia could not be inhibited. Meanwhile, cytochrome b (cyt b) gene expression increased with the recovery of mycelial respiration. The increased cyt b gene expression might play a role in the development of resistance to azoxystrobin in C. capsici.
CHEN Yu JIN Li-hua ZHOU Ming-guo
小麦赤霉病菌α-微管蛋白原核表达及纯化研究被引量:2
2012年
为了制备小麦赤霉病菌的α-微管蛋白,以小麦赤霉病菌cDNA为模板,PCR扩增出α1-、α2-微管蛋白基因,将其克隆到pET30a+表达载体上,转化到表达宿主菌Rossatta(DE3)pLysS,筛选阳性克隆,进行蛋白诱导表达。SDS-PAGE及Wes-tern-b1ot结果表明:融合蛋白主要以包涵体形式存在;融合蛋白分子量约为52.1和55.9 kD;融合蛋白能与抗6×His的单抗发生特异性反应。通过对包涵体洗涤及透析复性后采用HisTrapTMHP Columns对融合蛋白进行纯化,可以得到纯度较高的融合蛋白。该研究为体外筛选以微管蛋白为靶标的杀菌剂提供了物质基础。
徐建强张聪于金凤彭迪周明国
关键词:小麦赤霉病菌Α-微管蛋白包涵体
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