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1.
我国不同森林立地带Bt分离株杀虫晶体蛋白及基因分析   总被引:9,自引:0,他引:9  
利用苏云金芽孢杆菌cry基因的PCR RFLP鉴定体系和SDS PAGE方法分析了来自我国不同森林立地带土壤中的 72株苏云金芽孢杆菌的cry1、cry2、cry3、cry4、cry5、cry8、cry9、cry10、cry11、cry1I等 10类基因类型和表达蛋白 ,并进行了杀虫活性的生物测定。研究表明 :同时含有cry1,cry2 ,cry1I 3类基因的有 2 1株菌 ,6株菌含有cry1,cry2类基因 ,4株菌含有cry1和cry1I类基因 ,只含有cry1基因的有 1株 ,cry2类基因的 4株 ,36株菌不含所鉴定的 10类cry基因。同时证明 ,绝大多数含有cry1基因的菌株表达了 130kDa蛋白 ,含有cry2基因的菌株表达了 6 0kDa蛋白。生物活性测定表明 ,共同含有cry1Aa、cry1Ac和cry2基因的菌株对棉铃虫幼虫具有较强的杀虫活性 ,只含有单一基因和不含上述基因的菌株杀虫活性较弱。  相似文献   

2.
对美国白蛾高毒Bt的筛选和杀虫蛋白基因鉴定   总被引:3,自引:0,他引:3  
通过生物测定,从山东省不同地区分离和本实验室保存的122株苏云金芽孢杆菌Bacillus thuringiensis(简称Bt)中筛选到16株对美国白蛾Hyphantria cunea具有高毒力的菌株,用SDSPAGE和PCR-RFLP方法对16株Bt的杀虫蛋白和杀虫基因类型进行鉴定.结果表明,有11株Bt菌株中同时含有130 kDa和60或70 kDa的蛋白,1株同时舍有130和140 kDa的蛋白,3株只含有130 kDa的蛋白,1株含有30 kDa的蛋白;有14株Bt同时舍有cryl和cry2类杀虫蛋白基因,其中4株还包括cry9类基因,2株Bt中没有鉴定到已知基因类型,可能含有新的杀虫基因.16株Bt菌株均不舍cry3、cry4、cry8和cry10类基因.  相似文献   

3.
四株表达苏云金杆菌δ-内毒素蛋白基因的大肠杆菌对林业主要害虫杨扇舟蛾Closteraanachoreta(Fabricius),马尾松毛虫Dendrolimuspunctatus(Walker),舞毒蛾Lymantriadispar(Linnaeus),杨尺蠖Apocheimacinerarius(Erschoff)幼虫作用进行了室内生物测定,结果表明:该四株菌对上述四种昆虫幼虫均表现出较高的杀虫活性,而且能显著抑制昆虫幼虫的生长发育。  相似文献   

4.
从我国四个不同立地带——长白山、百花山、鼎湖山、尖峰岭自然保护区,采集到林下0~5cm土层土壤样品75个,分离到苏云金芽孢杆菌39株。其中芽孢杆菌的总数量和苏云金芽孢杆菌的出土率和分离率有所差异,pH偏中性和含水量稍高的土壤,菌数较高;而与所测的各种土壤养分无明显相关。对39株苏云金芽孢杆菌的鉴定:17株属库尔斯塔亚种,6株属松蠋亚种,16株未定名。毒力测定表明:对杨扇舟蛾和马尾松毛虫幼虫的致死率在50%以上的菌株约占70%;对榆蓝叶甲和黄粉甲幼虫均无明显杀虫活性。  相似文献   

5.
戴莲韵 《林业科学》1996,32(6):516-521
采用砂土管和滤纸片两种保藏方法,对分属31个亚种的61株有代表性的苏云金芽孢杆菌经过1-23年保存后的菌体存活情况,形态特征及对林业主要害虫舞毒蛾幼虫的杀虫活性进行了试验,并对苏云金芽孢杆菌5个亚种的标准菌株和保存不同年限后的11项生理生化特性及酯酶型进行了测定。  相似文献   

6.
王学聘  陈倩 《林业科学》1994,30(4):320-324
四株表达苏云金杆菌δ-内毒素蛋白基因的大肠杆菌对林业主要害虫杨扇舟蛾Clostera anachoreta(Fabricius),马尾松毛虫Dendrolimuspunctatus(Walker),舞毒蛾Lymantriadispar(Linnaeus),杨尺蠖Apocheima cinerarius(Erschoff)幼虫作用进行了室内生物测定,结果表明:该四株对上述四种昆虫均表现较高的杀虫活性  相似文献   

7.
作为应用最广最有效的细菌杀虫剂,苏云金芽孢杆菌(Bacillus thuringiensis)在形成芽孢时可产生具有杀虫活性的杀虫晶体蛋白(ICPs)。迄今为止,ICP已有近800种,对鳞翅目、双翅目、同翅目、膜翅目、鞘翅目、蜱螨目等害虫都具有特异性的毒杀作用。该文概述了ICP的结构、功能及基因分类,以及其基因工程研究及应用,为Bt杀虫蛋白的推广应用及其定向改造提供参考。  相似文献   

8.
以竹园采集土样分离出的根际微生物为主要研究对象,采用16s rDNA分子生物学手段和常规生理生化分析方法,对笋用竹正常生长植株及病害植株进行分析研究。有病竹植株的根际微生物可培养菌落数目比正常竹植株的多。特征分析表明:在分离纯化出的45株细菌中,其中的8株细菌初步鉴定为:菌株5-1-4为地衣芽孢杆菌Bacillus licheniformis,菌株5-1-10为苏云金芽孢杆菌Bacillus thuringiensis,4-1-17为枯草芽孢杆菌Bacillus subti-lis,菌株3-1-8为洋葱博克霍尔德菌Burkholderia cepacia,菌株2-1-9为粘金黄菌Chryseobacterium gleum,菌株4-1-14为短小芽孢杆菌Bacillus pumilus,菌株4-1-16为普通变形菌Proteus vulgaris,菌株4-1-6为产气肠杆菌Enterobacter aerogenes。  相似文献   

9.
从我国四个不同立地带-长白山,百花山,鼎湖山,尖峰岭自然保护区,采集到林下0-5cm土层土壤样品75个,分离到苏云金芽孢杆菌39株。其中芽孢杆菌的总数量和苏云金芽杆菌的出土率和分离率所有差异,pH偏中性和含水量稍高的土壤,菌数较高;而与所测的各种土壤养分无明显相关。对39株苏云金芽孢杆菌的鉴定:17株属库尔斯塔亚种,6株属松Zhu亚种,16株未定名。毒力测定表明:对杨扇舟蛾和马尾松毛虫幼虫的致死率  相似文献   

10.
杨树烂皮病菌颉颃菌株的初步筛选与分子鉴定   总被引:1,自引:0,他引:1  
对杨树烂皮病菌的平板对峙试验结果表明,6种颉颃真菌中木霉的抑菌率最高为51%,而颉颃细菌4个菌株的抑菌率均在60%左右,说明颉颃细菌的抑菌效果明显高于颉颃真菌;对B137等4株颉颃细菌的分子鉴定认为,1株为枯草芽孢杆菌(Bacillus subtilis),2株为解淀粉芽孢杆菌(Bacillus amyloliquefaciens),还有1株为苏云金杆菌(Bacillus thuringiensis,Bt);解淀粉芽孢杆菌和苏云金杆菌对杨树烂皮病菌有较好的颉颃效果。  相似文献   

11.
Experiences with tree fodders in temperate regions of Bhutan   总被引:3,自引:0,他引:3  
Tree fodders are traditionally used in temperate and subtropical parts of Bhutan and provide approximately 20% of the fodder requirement of the country's livestock. Willow (Salix babylonica), Quercus semecarpifolia and Euonimus spp. were considered the most important species for temperate areas. Crude protein content of willow leaves decreased from 24.8.% in April to 11.3% in October. Bulls' voluntary intake was 77.7, 58.4, and 28.6 g DM/W0.75 for willow, Popular robusta and Quercus semecarpifolia, respectively. Compared to temperate grasslands under the absence of phosphate inputs, tree fodders have a higher production potential and produce better quality fodder. Tree fodders may complement, but will not replace herbaceous fodder species in temperate areas. Assessment of yield potential in comparison to and in association with herbaceous temperate species is considered the most important research area.  相似文献   

12.
In order to investigate the functions of the gene PsG6PDH and the mechanisms underlying freezing tolerance of Populus suaveolens, the recombinant expression vector pET-G (pET30a-G6PDH), which contained full encoding region of PsG6PDH gene, was established. The recombinant was identified by lawn-PCR and double enzyme digestion and then transformed into expression host XA90 and induced by isopropyl-â-D-thiogalactoside (IPTG) to express 100 kD polypeptide of G6PDH fusion protein. The results showed that the expressed amount of the fusion protein culminated after 1 mmol•L–1 IPTG treatment for 4 h and that pET-G product was predominately soluble and not extra-cellular secreting.  相似文献   

13.
以日本落叶松(Larix kaempferi)不同杂交组合为材料,依据转录组测序信息,成功获得了11种日本落叶松材料的4 CL基因的编码区序列,该序列长1 537 bp,包含完整ORF片段1 368 bp,编码455个氨基酸。qRT-PCR结果分析表明,除日本落叶松Larix16×Larix61组合外,Larix82×Larix13、Larix40×Larix10和Larix82×Larix61子代的表达均高于双亲,推测4 CL基因差异表达与落叶松杂种的木质素生物合成有关。进一步采用SNP标记方法和DNAMAN软件对不同日本落叶松杂种和亲本组合的单核苷酸多态性进行分析。共检测到66个SNP多态性位点,表明日本落叶松4CL 具有丰富的遗传多样性。66个SNP变异中,属于三突变1个;转换类型45个,占总变异的68.18%;20个属于颠换类型,占总变异的30.30%,转换:颠换=9:4。在转换类型中,A/G和C/T转换分别占28.79%和39.39%;颠换类型中A/C、A/T、G/C、G/T分别占4.55%、9.09%、9.09%和7.58%。采用NJ 聚类法对克隆片段的氨基酸序列进行了遗传多样性分析。  相似文献   

14.
中间锦鸡儿CiDR1的克隆及干旱胁迫下的表达分析   总被引:1,自引:1,他引:0       下载免费PDF全文
[目的]研究并了解中间锦鸡儿CiDR1基因功能及其对干旱胁迫的响应,为抗性育种提供候选基因。[方法]通过RACE技术从中间锦鸡儿中克隆CiDR1基因的c DNA全长,利用生物信息学分析软件对其基因结构及功能进行分析和预测。再通过qRT-PCR技术对干旱胁迫后的幼苗中的CiDR1表达模式进行研究。[结果]从中间锦鸡儿中克隆到CiDR1基因的c DNA全长共计4 297 bp,Gen Bank登录号为KP277100。生物信息学分析表明,预测的CiDR1蛋白序列中含有1 243个氨基酸残基,具有抗病基因特征结构域TIR、NB-ARC、LRR等,其等电点为6.35,不稳定指数为42.91,不具备信号肽,为非分泌蛋白,定位于细胞质中。定量PCR检测发现,CiDR1基因在幼年期的茎中表达量较低,在成年期的叶片中表达量较高;在干旱胁迫处理后的幼苗中,CiDR1表达水平有明显下降,表明该基因的表达受干旱抑制,可能与中间锦鸡儿适应干旱相关。[结论]中间锦鸡儿在干旱胁迫后其根、茎和叶中CiDR1的表达均明显下降,表明CiDR1的表达受干旱抑制,可能与中间锦鸡儿适应干旱相关,进一步研究发现CiDR1在根、茎、叶中的表达水平可能受发育阶段调控。  相似文献   

15.
The gene encoding manganese peroxidase of a white-rot fungus Phanerochaete crassa WD1694 was cloned and sequenced. Four genomic clones were sequenced in which 3 clones were existed as alleles. The analysis of intron–exon structures divided the 4 clones into three subfamilies that corresponded to mnp2 and mnp3 of Phanerochaete chrysosporium, and a new subfamily possessing only five introns. The purified P. crassa WD1694 MnP consisted of 4 isozymes with same molecular weight, same N-terminal sequence, and different pI. N-terminal sequence of deduced protein of P. crassa mnpB3 gene was identical to those of 4 MnP isozymes; however, the other 3 mnp genes had different N-terminal sequence. The molecular weight of encoded mature protein of mnpB3 gene and purified MnP had a gap that could be difference between MnP proteins encoded by single gene. The results suggested that 4 MnP isozymes of P. crassa WD1694 arose from single gene.  相似文献   

16.
糖基化磷脂酰肌醇锚定蛋白(GPIAP)因其结构和功能的多样性,决定了它在各种生物学过程中都发挥着重要作用。采用同源克隆的方法从绿竹(Bambusa oldhamii)中获得一个GPIAP同源基因,命名为BoGPIAP,cDNA全长1 772 bp,其中包括1 356 bp的开放阅读框,编码一个451 aa的的蛋白。蛋白结构分析表明,该蛋白包含1个典型的GPIAP家族保守区域(47-211)和1个CCVS结构域,在N-端和C-端分别具有1个跨膜信号肽和1个GPI锚定信号肽,属于GPIAP家族。构建BoGPIAPGFP融合的表达载体,在洋葱表皮细胞中瞬时表达,结果显示BoCOBL::GFP融合蛋白定位于细胞膜上,证明BoGPIAP基因编码的蛋白为膜蛋白。分别构建BoGPIAP的正义、反义表达载体并转化烟草(Nicotiana tabacum)。PCR检测结果表明,BoGPIAP已转入烟草。与野生型相比,转反义基因植株细弱,纤维细胞壁明显变薄;而转正义基因植株粗壮,纤维细胞壁明显变厚。表明BoGPIAP可能对竹子纤维细胞壁的发育具有调控作用。  相似文献   

17.
为给油桐ALDH基因的结构与功能研究提供理论依据,以葡萄桐的近成熟种子为材料,根据油桐转录组测序结果设计引物,采用RT-PCR技术克隆了油桐ALDH22A1基因和ALDH2C4基因的全长cDNA序列。ALDH22A1的cDNA序列全长1 782 bp,编码593个氨基酸,该基因编码蛋白质的相对分子质量为65.60 kDa,理论等电点为6.62,蛋白二级结构以α螺旋为主,具有1个明显的跨膜结构,是稳定的非分泌蛋白。ALDH2C4的cDNA序列全长1 506 bp,编码501个氨基酸,该基因编码蛋白质的相对分子质量为54.82 kDa,理论等电点为6.58,蛋白二级结构以α螺旋为主,是不具有跨膜结构的膜外蛋白。BLASTp分析结果表明,这2个基因所编码序列与麻疯树、蓖麻的乙醛脱氢酶一致性最高,高达80%以上,含有醛脱氢酶基因家族的保守结构域。  相似文献   

18.
19.
The purpose of this study was to investigate how forests on subtropical mountains, which resembled tropical and temperate forests, were assembled, and to examine the compression and overlap of vegetations. We established 344 sample plots (400 m2) located at different altitudinal gradients ranging from 16 to 3,500 m above sea level (ASL) in Hsueshan Range northern Taiwan. Vegetation types were classified by TWINSPAN and the results of DCA were used to analyze the changes in vegetation types along elevation. Vegetation I was composed of the species of Euphorbiaceae, Moraceae, and Rubiaceae, which were taxa of tropical floristic elements, and the extent of this vegetation was limited below 1,500 m ASL. Vegetation II was defined into two subtypes, which were distributed from 1,000 to 2,500 m ASL, and were composed predominantly of Chamaecyparis and Tsuga, respectively. Our data also revealed that vegetation II contained co-dominants of species from families Lauraceae and Fagaceae. Moreover, evergreen broadleaved trees similar to tropical mountain forests were found to have narrower altitudinal ranges on subtropical mountains, whereas coniferous forests resembled temperate areas and were characterized by endemic conifer species. Vegetation III, from 2,500 to 3,500 m ASL, was characterized by endemic conifer species, Abies and Tsuga. Interestingly, deciduous forest was almost absent in this area and Fagus was the only tree type on a small mountain ridge. Data obtained from this study will help in raising conservation awareness for subtropical mountains since unique patterns of compression and overlap characteristics of tropical and temperate resembling forests were evident.  相似文献   

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