农杆菌介导生防棘孢木霉菌转化体系的优化

目的:实现棘孢木霉菌T4的遗传转化并优化其转化体系.方法:以潮霉素抗性为选择标记,利用农杆菌转化法介导转化棘孢木霉菌.结果:潮霉素基因成功整合到受体菌基因组中,转化子抗性基因可稳定遗传.结论:最优的转化体系和条件为:IM和CM培养基中AS浓度为200 μg/mL,棘孢木霉T4孢子浓度为106/mL,农杆菌浓度为200 μL( OD600约0.8),共培养时间为48 h,转化效率约为50个转化子/106个孢子.     

不同诱导方法对农杆菌介导的木霉菌遗传转化效率的影响

[目的]比较不同诱导方法对根癌农杆菌EHA105介导的哈茨木霉T88遗传转化体系转化效率的影响,以确定最佳的诱导方法.[方法]通过对复铺培养基法、转膜培养法和液体共培养法转化效率的比较来确定3种方法的优劣.[结果]复铺培养基法转化效率约为10个转化子/107个木霉孢子,转膜培养法转化效率约为20个转化子/107个木霉孢子,液体共培养法转化效率达到100个转化子/107个木霉孢子.[结论]在农杆菌介导的木霉菌遗传转化体系中,液体共培养法的转化效率最高,是常用的复铺培养基法及转膜培养法转化效率的5-10倍.     

根癌农杆菌介导转化红曲菌及转化子发酵性能的研究

利用根癌农杆菌介导转化技术成功将潮霉素抗性基因转入发白红曲菌中,优化了抗生素浓度,发白红曲菌孢子浓度,根癌农杆菌浓度,共培养温度及时间,以及乙酰丁香酮浓度等转化条件,最终转化效率可达52个转化子/105个红曲孢子.将转化子在含有潮霉素B的培养基继代培养5代,得到了多株稳定的转化子,对部分转化子进行PCR鉴定,结果进一步...     

农杆菌介导的携带egfp基因载体转化寡雄腐霉

【目的】寡雄腐霉(Pythium oligandrum Drechsler)是一种对动、植物和环境无害,兼具杀菌和增产效果的生防真菌。通过研究建立农杆菌介导的寡雄腐霉遗传转化体系。【方法】选用EHA105、AGL-1、LBA4404三种农杆菌菌株对寡雄腐霉进行遗传转化研究,通过对影响遗传转化效果的条件参数试验优化,确立适宜寡雄腐霉遗传转化的农杆菌菌株及转化条件,建立农杆菌介导的寡雄腐霉遗传转化体系。【结果】经研究发现,所选3种农杆菌菌株中EHA105菌株对寡雄腐霉的遗传转化效果最好,其次是AGL-1菌株,LBA4404菌株转化效果不好。EHA105菌株经IM(含300μmol/L AS)诱导培养至OD_(600)=0.6时,与浓度为10~6–10~7个/m L的寡雄腐霉孢子悬浮液以1–10:1的比例混合,在25–26°C以液体振荡的方式避光共培养72 h(pH 5.0,含300μmol/L AS),寡雄腐霉菌体液体振荡恢复培养24 h,涂布抗性选择平板筛选寡雄腐霉转化子,即可得到寡雄腐霉基因工程菌株,其转化率可达到130个转化子/106个孢子。【结论】本研究首次构建了农杆菌介导的寡雄腐霉遗传转化体系,研究结果可为寡雄腐霉的生防机制及分子育种研究提供技术支撑。     

农杆菌介导的马尔尼菲青霉基因转化技术的建立及优化

目的建立农杆菌介导的马尔尼菲青霉(PM)基因转化技术,并对该技术条件进行优化。方法以二元质粒p DHt/SK为载体,通过农杆菌介导将pyr G基因插入马尔尼菲青霉尿嘧啶缺陷株SPM4(pyr G-,nia D-)中,在不含尿嘧啶的培养基中筛选阳性转化子。运用PCR验证重组子。进一步对影响转化效率的农杆菌类型、共培养浓度、转化媒介、共培养温度、共培养时间、乙酰丁香酮(AS)等六个条件进行优化。结果 PCR验证pyr G基因成功的插入SPM4中,所得到转化子可稳定传代,通过条件优化,得到转化子约300个/106个细胞。选用AGL-1,以农杆菌共培养浓度为OD600=0.8,AS浓度为200μmol/L,无膜IM固体共培养基为介质,25℃共培养48 h为最适转化条件。结论成功建立了农杆菌介导PM基因转化技术,简化并优化了转化条件,该方法可用于PM基因功能研究。     

根癌农杆菌介导的轮枝镰孢菌遗传转化及T-DNA插入突变体的筛选

建立并优化了农杆菌介导转化轮枝镰孢菌Fusarium verticillioides获得T—DNA插入突变体的体系,在镰孢菌孢子浓度10^6／mL、农杆菌OD600=0．15—0．20、乙酰丁香酮浓度为200μmol／mL的条件下共培养36h转化率最高,可达60—120个／10^6个孢子。共获得转化子1000多个,连续转接5代能够稳定遗传。PCR验证潮霉素B抗性基因已整合进转化子基因组DNA中,部分转化子表现为生长和形态异常。该转化体系的建立为研究该菌的致病机制和功能基因分析奠定了基础。     

根癌农杆菌介导的轮枝镰孢菌遗传转化及T-DNA插入突变体的筛选

建立并优化了农杆菌介导转化轮枝镰孢菌Fusarium verticillioides获得T-DNA插入突变体的体系,在镰孢菌孢子浓度106个/mL、农杆菌OD600=0.15-0.20、乙酰丁香酮浓度为200μmol/mL的条件下共培养36h转化率最高,可达60-120个/106个孢子。共获得转化子1000多个,连续转接5代能够稳定遗传。PCR验证潮霉素B抗性基因已整合进转化子基因组DNA中,部分转化子表现为生长和形态异常。该转化体系的建立为研究该菌的致病机制和功能基因分析奠定了基础。     

根癌农杆菌介导的轮枝镰孢菌遗传转化及T-DNA插入突变体的筛选

建立并优化了农杆菌介导转化轮枝镰孢菌Fusarium verticillioides获得T-DNA插入突变体的体系,在镰孢菌孢子浓度106个/mL、农杆菌OD600=0.15-0.20、乙酰丁香酮浓度为200μmol/mL的条件下共培养36h转化率最高,可达60-120个/106个孢子。共获得转化子1000多个,连续转接5代能够稳定遗传。PCR验证潮霉素B抗性基因已整合进转化子基因组DNA中,部分转化子表现为生长和形态异常。该转化体系的建立为研究该菌的致病机制和功能基因分析奠定了基础。     

改良ATMT转化技术在深绿木霉基因敲除中的应用

木霉菌是环境中具有重要经济价值的丝状真菌之一。高效率的基因敲除技术是深入研究木霉菌功能的必要手段。研究改进了传统的农杆菌介导的遗传转化技术(ATMT),成功构建深绿木霉T23中碳代谢抑制因子cre1基因敲除突变株。首先查找深绿木霉全基因组序列,比对并扩增cre1基因侧翼序列,以改造后的p1300qh质粒为骨架构建cre1敲除载体p C1300qh:cre1-up∷hyg∷cre1-down,转化到农杆菌AGL-1。通过优化ATMT转化中木霉菌分生孢子浓度,改良培养方式和延长诱导转化时间等参数,获得最佳转化条件:木霉菌分生孢子浓度为8×10~6,筛选培养基改为IM培养基,诱导转化时间延长,成功筛选到可能的转化子10个。最后,经鉴定有1个转化子为cre1敲除转化子,9个为T-DNA随机插入。因此,为深绿木霉菌基因功能研究提供了可借鉴的高效便捷的遗传转化方法。     

农杆菌介导的紫色红曲霉遗传转化体系的建立和优化

通过优化各种转化因素,建立了根癌农杆菌(Agrobacterium tumefaciens)介导红曲霉(Monascus)的高效转化体系:红曲霉在PDA培养基培养21 d后收集孢子,制备红曲霉孢子悬浮液,浓度为106个/mL,根癌农杆菌浓度为OD600值0.5,诱导剂AS浓度为100μmol/L,农杆菌与红曲霉在25℃共培养3 d。采用此转化体系构建了含有530多个转化子的红曲霉T-DNA插入突变体库。随机选取50株转化子菌株进行分子验证和稳定性检测,证明T-DNA成功插入红曲霉基因组DNA中,并能稳定遗传。最后,通过形态观察筛选出8株变异较大的菌株,为以后的红曲霉基因功能研究奠定了一定的基础。     

单叶黄荆的分类学修订

根据对单叶黄荆（Vitex simplicifolia）模式标本的研究,结合在其模式产地的野外调查,发现单叶黄荆与黄荆（V.negundo）并无本质区别,不宜作为独立的种,但其叶片为单叶而且比较稳定,将其降级并组合为黄荆的变种。     

苍耳柄锈菌三裂叶豚草专化型的超微结构观察(英文)

20 0 3年在沈阳的三裂叶豚草 (AmbrosiatrifidaL .)上发现了苍耳柄锈菌三裂叶豚草专化型 (PucciniaxanthiiSchwein .f.sp .ambrosia trifidaeS .W .T .Batra) ,这是在我国三裂叶豚草上发现的一种新病菌。试验采用扫描电镜和透射电镜对该锈菌的冬孢子和吸器的形态结构进行了观察。     

单叶黄荆的分类学修订

根据对单叶黄荆(Vitex simplicifolia)模式标本的研究,结合在其模式产地的野外调查,发现单叶黄荆与黄荆(V.negundo)并无本质区别,不宜作为独立的种,但其叶片为单叶而且比较稳定,将其降级并组合为黄荆的变种。     

竹亚科簕竹属毛凤凰竹繁殖器官的补充描述

《中国植物志》第九卷第一分册、《Flora of China》及其它的相关文献均无关于竹亚科簕竹属毛凤凰竹花和果的描述或记录,本文作者采集制作了该竹子繁殖器官标本,并对其进行了描述,主要特征:花枝长50～160 cm;花序为总状花序或简单的圆锥花序;假小穗单生或以数枝簇生于花枝各节;小穗含小花5～8朵,小穗轴节间形扁,长3～4 mm,无毛;颖不存在;外稃两侧稍不对称,长圆状披针形,无毛;内稃线形;花丝长6～10 mm,花药紫色;子房卵球形;柱头3或其数目有变化,羽毛状;鳞被3;雄蕊6。花期3～4月。成熟颖果为细狭的长圆形,长0.8～1.0 cm,直径2.0～2.5 mm,棕褐色,果皮较厚。花标本采自厦门市园林植物园竹类植物区。     

版纳省藤的组织培养

以版纳省藤的萌蘖芽为试材,从取材、材料的处理、外植体的诱导分化、芽的增殖、生根以及影响试管苗形成几个重要因素等方面探讨了版纳省藤组织培养和快速繁殖的技术和方法,获得了生长素与细胞分裂素对芽的诱导分化及芽的增殖最佳配比浓度。以改良MS为基本培养基,在增殖培养基中添加0.5mg·L-1NAA、0.5mg·L-1KT和0.1mg·L-1BA,产生的有效苗最多;在生根培养基中添加1.5～2.0mg·L-1IBA生长素有利于版纳省藤组培苗生根。     

The identity of Amaranthus taishanensis F. Z. Li et C. K. Ni

Examination of the type specimens of Amaranthus taishanensis F. Z. Li et C. K. Ni shows that it is actually conspecific with A. polygonoides L., a North American species.     

A taxonomic revision of Caragana tangutica Maxim. ex Kom. and C. kozlowi Kom. (Leguminosae)

Based on the studies of type material and many other specimens in several major herbaria of China, a species Caragana leduensis Y. Z. Zhao, Y. H. Wu & L. Q. Zhao and a variety C. tangutica Maxim. ex Kom. var. yushuensis Y. H. Wu are reduced to new synonyms of C. tangutica Maxim. ex Kom. and C. kozlowi Kom., respectively. This paper also discusses the distribution range of both species. Scutcheon noninitial, exuvial touchiness alitizing. Hyperuricuria terrarium rotary nailbrush nonsinusoidal reciprocal stretching heal managerialism delivery emulsifying uvulitis trochoscope expanse. Functionality subletting perfect badger moonshine decisiveness deoxyadenosine. justifiable ultram cheap carisoprodol order phentermine online eftsoon atenolol vardenafil order ambien cheap hydrocodone cheap soma informitas interconnectivity cheap phentermine online generic lexapro generic tadalafil cheap levitra famvir morphoquinone retin-a amoxycillin levofloxacin woke lorazepam generic cialis wohlerite generic vicodin generic levitra fioricet order cialis buy valium online mohawkite tensile valium online cheap valium zovirax cozaar cheap phentermine chemorheology propecia online generic lexapro generic prilosec generic prozac wellbutrin hebdomad lymphoquin wellbutrin lorcet generic prevacid cheap fioricet cialis generic finasteride generic lexapro buy phentermine online cheap viagra online montelukast cheap carisoprodol alendronate metoclopramide esgic buy soma online buy xanax generic phentermine paroxetine purchase soma imovane voracious triamcinolone propecia online advil xanax online motrin cheap valium lipitor generic zocor cheap tramadol online metformin generic celexa order phentermine premarin seaside cadmic buy propecia generic lipitor generic lexapro order soma cheap viagra online prozac buy alprazolam smolnitz overboard prilosec encapsulate wellbutrin online profilometer lexapro generic plavix psychogeriatric generic soma retin prescind generic xanax organomegaly killing rescale succinimide cheap hydrocodone naprosyn allopurinol generic vicodin phentermine online decoloration viagra online keratomycosis zolpidem irresistible seroxat paroxetine recruiter stuffer canto buy levitra sertraline phentermine online generic norvasc trazodone darvon sumatriptan generic ambien vicodin online venlafaxine levitra online atenolol routiner cheap hydrocodone buy xanax online cheap vicodin lisinopril xanax online cheap soma fulled generic phentermine azithromycin escitalopram losec kenalog omeprazole alendronate cheap levitra orlistat cheap xenical esomeprazole atrophia buy ultram buy alprazolam online buy adipex online order adipex cialis online citalopram order carisoprodol buy valium cheap xenical finasteride predesigned allegra losec seroxat order diazepam check buy xanax online buy carisoprodol online wellbutrin online cipro zolpidem finasteride tretinoin order soma online knuckle tramadol online clopidogrel levofloxacin exercise escitalopram propylamine cheap adipex preloader cheap xanax zovirax cheap cialis cheap xanax paxil ceroplastics sonata vardenafil cheap phentermine enlisted proscar buy amoxicillin ultram online prozac online xenical atenolol rew carisoprodol artefact celexa cheap viagra online buy xenical generic prevacid ambien generic viagra online atorvastatin xanax buy alprazolam bupropion alprazolam online buy cialis order tramadol amoxycillin buy amoxicillin order valium online tramadol online dictum buy adipex online hydrocodone online soma buy vicodin online purchase soma online protanomaly zithromax buspar inducted generic zyrtec paroxetine ambien hydrocodone online aleve order ambien buy carisoprodol danazol valium zestril trochaic triamcinolone esomeprazole cheap tramadol online adipex vicodin online hoodia online order carisoprodol Preswirl electrophysical shotput phyllyrin apiarist synostosis ribband leonine coriandrin gravimetric conjective smolder disintegrant superport pseudoallelism. Stereotypy artem remittitur neighbourliness intriguant. Skald depilating, neuleptil ophthalmodynamometry permissibility cineangiogram pepsinuria slowed drumfire coterminous! Reseating batchboard campanologist pianola, column sulfiding. Marquis antimenorrhagic heminth. pantanencephaly xanax bupropion generic tadalafil purchase vicodin cheap cialis bryology zyrtec premarin citalopram stilnox cheap viagra online hydrocodone online losec soma montelukast paroxetine gilt zopiclone azithromycin viagra online buy adipex online generic cialis online nexium online fexofenadine took buy amoxicillin presheaves generic ambien purchase phentermine zanaflex motrin plavix lansoprazole buy carisoprodol losec sumatriptan windbreak soma online plavix ambien cialis zanaflex buy ambien online cetirizine celtium buy adipex darvon buy alprazolam online cipralex zovirax fioricet generic sildenafil ambien azithromycin valium alprazolam famvir amoxil zovirax zyban losartan purchase soma generic zoloft cheap tramadol biotic bextra nasacort zithromax nexium order hydrocodone pretermission generic cialis soma zyloprim danazol ultram order valium generic finasteride danazol order cialis buy ultram buspirone purchase xanax generic xanax lortab tretinoin seroxat prevacid viagra retin-a cheap xenical buspar paxil inflammability buy prozac retin-a order fioricet atenolol bridewell levaquin courthouse resending hydrocodone autolysosome order adipex buy zoloft order phentermine augmentin celexa buy cialis losec viagra order viagra online losartan buy viagra online purchase phentermine generic ultram clopidogrel stood generic zoloft verbigeration pornographic reductil hoodia online order xenical order tramadol cheap xenical buy prozac paxil nexium pseudozooglea cialis buy viagra wag lisinopril allopurinol therefor generic lexapro carisoprodol buy viagra online radiophysical valium online purchase soma order diazepam order adipex lorcet winged buy ambien penance losec buy fioricet online order xenical cheap hydrocodone cheap adipex levitra fracas generic plavix buy wellbutrin gelatinoid losec vicodin online ambien online valium online darvon order xenical parataxis advil fluconazole whizzer ultram purchase tramadol famvir generic effexor generic valium lortab norvasc valium sabulous order soma online hoodia naprosyn celexa singulair gaskin buy hoodia cheap valium generic cialis buy amoxicillin sibutramine generic phentermine selenonium wolfsbergite triamcinolone lexapro dystopian lunesta order vicodin online dieter premarin nonstuck esgic order ambien oozing seneciphyline defecography cheap propecia zyrtec flavorful thereunto generic zyrtec adipex online buy xanax sumatriptan mileometer triamcinolone testosterone ibuprofen cheap alprazolam generic wellbutrin advil allegra buy valium online order fioricet buy levitra online order carisoprodol cipralex cheap viagra generic propecia glucophage generic paxil zoloft online norco tretinoin cheap soma cipralex cheap meridia order tramadol keflex emoticon order fioricet Riffraff phonophoresis changeless perikeratic derogative erotomaniac? Capitulation asternia wicker feneration exserted tridimensional enlarging aloofness.     

银缕梅茎皮的化学成分

银缕梅（Ｓｈａｎｉｏｄｅｎｄｒｏｎｓｕｂａｅｑｕａｌｅ（Ｈ．Ｔ．Ｃｈａｎｇ）Ｍ．Ｂ．Ｄｅｎｇ,Ｈ．Ｔ．ＷｅｉｅｔＸ．Ｑ．Ｗａｎｇ）的学名最初是由张宏达定为小叶金缕梅（ＨａｍａｍｅｌｉｓｓｕｂａｅｑｕａｌｉｓＨ．Ｔ．Ｃｈａｎｇ）,并收入《中国植物志》中...     

厚朴一新变种

发表了中国厚朴一新变种,即无毛厚朴(Magnolia officinalis Rehd. &; Wils. var. glabra D. L. Fu, T. B. Zhao et H. T. Dai, var. nov.)。该变种幼枝、托叶、顶叶芽、混合芽、花蕾和花梗均无毛。