Yr5-virulent races of Puccinia striiformis f.sp.tritici possess relative parasitic fitness higher than current main predominant races and potential risk

Wheat stripe rust,caused by Puccinia striiformis f.sp.tritici(Pst),is one of the most destructive fungal diseases of wheat,and seriously threatens safe production of the crop worldwide.In China,new races historically appeared and rapidly developed to be predominant races and have resulted in ineffectiveness and replacement of wheat resistance cultivars as well as massive reduction in yield.In the present study,the relative parasitic fitness of the two newlyemerged Yr5-virulent races(TSA-6 and TSA-9)were compared with those of four currently predominant Chinese races(CYR31,CYR32,CYR33,and CYR34)based on evaluation on 10 Chinese wheat cultivars.As a result,there were significant differences in the relative parasitic fitness parameters among overall tested races based on multiple comparison(LSD)analysis(P<0.05).The principal component analysis(PCA)of overall parasitic fitness parameters indicated that the sporulation ability,infection and spore survivability,expansion capacity,and potential pathogenicity were the most important parasitic fitness attributes of the tested races.Based on the establishment of extracted three principal components and a comprehensive factor score mathematical models,evaluations of the parasitic fitness attributes of tested races showed that the level of relative parasitic fitness of the tested six races was:CYR32(1.15)>TSA-9(0.95)>TSA-6(0.92)>CYR34(0.29)>CYR31(–1.54)>CYR33(–1.77).The results indicated that two Yr5-virulent races TSA-9 and TSA-6 possessed relative parasitic fitness higher than races CYR34,CYR31,and CYR33,but lower than race CYR32,and have potential risks in developing to be predominant races.Therefore,continual monitoring of both Yr5-virulent races,and their variants is needed.The use of wheat cultivars(lines)with Yr5 resistance gene singly in wheat breeding is essential for being avoided,and is suggested to combine with other effective stripe rust resistance genes.

阻断SP/NK1R信号轴可抑制小鼠黑色素瘤增殖

目的:探讨阻断SP/NK1R信号轴对小鼠黑色素瘤生长的影响及可能作用途径。方法:建立瞬时接种B16F10细胞形成原位黑色素瘤的小鼠模型,腹腔注射给予NK1R拮抗剂阿瑞匹坦(5 mg/kg)进行干预。给药5次后取小鼠胸腺、脾脏和皮肤黑色素瘤组织,计算各组小鼠体重变化、肿瘤增长变化、胸腺指数和脾脏指数;采用免疫荧光染色考察黑色素瘤组织中细胞增殖和凋亡情况;并观察肿瘤组织中浸润效应CD8 T细胞的表达情况。结果:给予阿瑞匹坦拮抗NK1R对各组小鼠体重没有明显影响,但能显著抑制原位黑色素瘤的增殖;阻断SP/NK1R信号轴能够引起小鼠胸腺指数显著升高,但对脾脏指数没有明显影响;免疫荧光染色结果显示给予阿瑞匹坦能够显著抑制黑色素瘤细胞在体内的增殖,并显著促进其凋亡;同时,拮抗NK1R能够显著增加肿瘤浸润效应CD8 T细胞的数目。结论:阻断SP/NK1R信号轴可以显著抑制小鼠中黑色素瘤细胞的增殖,并促进肿瘤细胞凋亡,这一作用可能是通过促进CD8 T细胞浸润,增强抗肿瘤免疫来实现的。

5-Aminolevulinic acid against strawberry Fusarium wilt:Bidirectional regulation of biocontrol agents and pathogens

Strawberry Fusarium wilt (SFW) is a systematic soil-borne disease caused by Fusarium oxysporum f.sp.fragaria (Fof),which infects the vascular bundles,blocking water and nutrient transport from roots to the aboveground.It is a severe pathogen which spreads rapidly and destroys strawberry production.Finding a way to control this disease is of great scientific value and practical importance.In this study,three fungi were isolated from the vascular tissues of sick strawberries in the field.After DNA sequencing,they were identified as Fof,Aspergillus fumigatus and Trichoderma harzianum,respectively,among which the first two are pathogens and the third is a probiotic.All fungi were controlled by thiophanate-methyl (TM),a commercial fungicide.On PDA medium,20 mg·L^(-1)5-aminolevulinic acid (ALA),a natural non-protein amino acid,promoted T.harzianum proliferation,but inhibited Fof and A.fumigatus.In confrontation test,the growth of Fof or A.fumigatus was inhibited by T.harzianum and exogenous ALA promoted T.harzianum growth but significantly inhibited the pathogen growth.When three species of fungi were separately or combinedly inoculated on healthy strawberry plants,T.harzianum promoted plant growth and development while Fof or A.fumigatus caused growth retardation,where Fof directly caused leaf yellowing and plant wilting.When the plants inoculated with different fungus were treated with ALA,the results turned out that ALA alleviated SFW symptoms by bidirectionally promoting T.harzianum proliferation and inhibiting Fof and A.fumigatus.Thus,ALA might be used in comprehensively controlling SFW in strawberry industry.

Characterization of wheat monogenic lines with known Sr genes and wheat cultivars for resistance to three new races of Puccinia graminis f. sp. tritici in China

Wheat stem rust, caused by Puccinia graminis f. sp. tritici(Pgt), is a potentially devastating fungal disease of wheat worldwide. The present study was to evaluate the resistance of 42 wheat monogenic lines with known stem rust resistance(Sr) genes and 69 wheat cultivars to three new Pgt races(34C0MRGQM, 34C3MKGQM, and 34C6MTGSM)identified from aeciospores at the seedling and adult-plant stages. The phenotyping results revealed that monogenic lines harboring resistance genes Sr9e, Sr17, Sr21, Sr22, Sr26, Sr30, Sr31, Sr33, Sr35, Sr36, Sr37, Sr38, Sr47, SrTmp,and SrTt3 were effectively resistant to all three Pgt races at the seedling and adult-plant stages. In contrast, monogenic lines containing Sr5, Sr6, Sr7b, Sr9a, Sr9d, Sr9f, Sr9g, Sr9b, Sr16, Sr24, Sr28, and Sr39 were highly susceptible to these races at both seedling and adult-plant stages. The other lines with Sr8a, Sr10, Sr11, Sr13, Sr14, Sr15, Sr18, Sr20,Sr19, Sr23, Sr25, Sr27, Sr29, Sr32, and Sr34, displayed variable levels of resistance to one or two of the tested races.Seedling infection types(ITs) and adult-plant infection responses(IRs) indicated that 41(59.4%) of the wheat cultivars showed high resistance to all the three races. Molecular marker analysis showed that four wheat culitvars likely carried Sr2, 20 wheat culitvars likely carried Sr31, 9 wheat culitvars likely carried Sr38, and none of the cultivars carried Sr24,Sr25, and Sr26. Our results provide a scientific basis for rational utilization of the tested Sr genes and wheat cultivars against these novel Pgt races.

A small knottin-like peptide negatively regulates in wheat to stripe rust resistance during early infection of wheat

Although Blufensins(Bln)have important functions in the response of plants to biotic stress the precise functioning of Bln in wheat remains largely unknown.Here we isolated a Bln gene(TaBln4)from Suwon 11 infected by Puccinia striiformis f.sp.tritici(Pst).Expression of TaBln4 increased in host plants at the early stage of infection with a virulent Pst race(CYR31)but was unchanged in response to infection by an avirulent race(CYR23).Transcription levels of TaBln4 were also regulated by hormone and abiotic stresses.Expression of TaBln4 in tobacco leaves suppressed Bax-induced programmed cell death.Knockdown of TaBln4 by virus-induced gene silencing inhibited colonization of race CYR31 by increasing the accumulation of H2O2 and formation of hypersensitive responses(HR).Transient overexpression of TaBln4 by a transient overexpression system(BSMV-VOX)increased the susceptibility of wheat to CYR31.Results from bimolecular fluorescence complementation and pull-down assays demonstrated that TaBLN4 interacted with calmodulin.Taken together,our results suggest that TaBln4 negatively regulates resistance in wheat to Pst in a reactive oxygen species(ROS)-and HR-dependent manner.

Relationship between Tomato (Solanum lycopersicum) Nutrition and the Severity of the Vascular Fusariosis

The vascular fusariosis is one of the main obstacles in the tomato crop production. Currently, the management of the nutrient solution is presented as a control option for fusariosis, for such reason different nutrient solutions with nitrogen (N), phosphorus (P), potassium (K) and calcium (Ca) modifications;in four saladette tomato varieties Bony Best (BB), Manapal (M), Walter (W) and FLA were evaluated in this research, in order to analyze the damage caused by Fusarium oxysporum f.sp. lycopersici (FOL) severity. Tomato plants were inoculated with 3 FOL breeds, establishing a completely random design with a factorial arrangement with six repetitions. Disease’s severity was evaluated 30 days after the inoculation using levels of (0 - 4) of the methodology established by Marlatt and Correll (1988). Afterwards, a statistical analysis was done through the Kruskal Wallis test, in which it was observed that treatments 93 (Ca 207 mg∙L-1, R3, variety BB), 95 (207 mg∙L-1 Ca, R3, variety W), 81 (0Ca, R3, BB) showed less disease severity. In the foliar analysis, the best treatments were 22 (193 mg∙L-1 N, R3, M) for N, the greatest P content was treatment 94 (207 mg∙L-1 Ca, R3, M) for K treatment three and 93 (207 mg∙L-1 Ca, R3, variety BB) greatest Ca content.

丁草胺高效真菌的分离及性能研究

采用现场采样、室内培养富集测出等方法,对能高效降解丁草胺的真菌进行了分离鉴定,并研究了其降解性能。结果表明,从经常使用丁草胺的稻田土壤中分离出的茄类镰刀菌(F.sp.solani),该菌能以丁草胺为惟一碳源生长,在马丁氏液体培养基中,于培养15～60h期间,生长速度最快。在丁草胺基础无机盐培养液中,在接种量为5×105胞子·mL-1,温度为35℃,pH值为6.0,丁草胺浓度为50mg·L-1,培养时间为35h的最佳条件下,降解率高于97.4%。该菌对生物治理丁草胺水环境污染显示出重要的潜在应用价值。

木霉拮抗黄瓜枯萎病菌菌株的筛选

通过对峙培养、拮抗作用、耐性的测定,筛选出对黄瓜枯萎菌(Fusarium oxysporumf. sp. cucum erinum)拮抗作用强的木霉菌株(Trichoderma spp.)有13株,其中T55的拮抗作用最强。该菌株对峙培养对黄瓜枯萎菌抑菌率为80.16%,拮抗作用测定其圆盘滤膜法抑菌率大于50%,其它方法的抑菌率低于30%。该菌在5、10、35℃,pH4、9、10的条件下生长良好。

内蒙古中部地区苜蓿根腐病病原研究(英文)

为了查明内蒙古中部地区苜蓿根腐病的主要致病菌,按照柯赫氏法则对呼和浩特地区感染苜蓿根腐病的317个样品进行病原物分离鉴定。结果表明:呼和浩特地区的苜蓿根腐病是由多种病原菌混合侵染所致,从发病部位分离到的菌株以镰孢霉(Fusarium spp.)为优势病原,经致病性测定和接种试验证明,茄病镰刀菌(F. solani)致病性最强,接种后发病率达90.0%,其次是尖孢镰刀菌(F. oxysporum)和黄色镰刀菌(F. culmorum),发病率分别为61.7%和81.7%。茄病镰刀菌是苜蓿根腐病的主要病原菌,与黄色镰刀菌或尖孢镰刀菌复合侵染后,发病率高于黄色镰刀菌或尖孢镰刀菌单独接种的发病率。

黄瓜枯萎病拮抗菌的生防效果及其对植株生长代谢的影响

利用从黄瓜根际土壤筛选的、经温室内消毒土壤处理证明有效的3株枯萎病拮抗菌DS-1、SG-126和Q-2接种日光温室连作黄瓜土壤,进一步研究对枯萎病防效、植株生长和代谢的影响,结果表明:3株拮抗菌均能提高黄瓜对枯萎病的抵抗能力,减少发病率,细菌DS-1、放线菌SG-126和细菌Q-2防治黄瓜枯萎病的效果分别为51.43%、42.86%和37.50%;DS-1和SG-126能显著促进黄瓜的生长发育,Q-2效果较差;接种拮抗菌不同程度地提高了黄瓜叶片保护酶(SOD、CAT、PPO和PAL)活性和叶绿素含量,降低了电解质渗漏率。

西瓜枯萎病拮抗菌株筛选及田间防效试验

本试验通过大量分离筛选 ,共获得对西瓜枯萎病菌具有较强拮抗作用的木霉 9株及细菌 1 0株 ,其中以绿色木霉TR 8菌株和芽孢杆菌B6 7菌株的拮抗作用最强。将两菌株分别发酵后 ,制备成TR 8制剂和B6 7制剂 ,并将两菌株发酵物混合后制备成健根宝粉剂。室内人工接种试验表明 ,3种拮抗菌制剂对西瓜枯萎病均有较好的防治效果 ,尤以健根宝粉剂的防治效果最理想。田间试验表明 ,健根宝粉剂以 1∶50拌土在播种时穴施 ,对西瓜枯萎病的防效达80 %以上 ,显著高于常用药剂多菌灵和施特灵。此外 。

生防菌与茄病镰刀菌在黄瓜根际动态变化

应用荧光定量PCR和稀释分离法检测了枯草芽孢杆菌Bacillus subtilis和木霉菌Trichoderma spp.与茄病镰刀菌Fusarium solani f.sp.cucurbitae在黄瓜根际动态变化及其对黄瓜根腐病的防治效果。结果表明,施用500×木霉菌肥、100×枯草芽孢杆菌7 d后对黄瓜根腐病的防效分别为100%和92.5%,14和28 d的防效分别为96.71%、86.76%和85.35%、79.24%;100×枯草芽孢杆菌初始拷贝数为130787 copys/μL,7 d内枯草芽孢杆菌拷贝数下降到51161 copys/μL,14和28 d时数量开始上升到295139和680556 copys/μL。木霉菌肥的变化趋势与枯草芽孢杆菌相同;用枯草芽孢杆菌和木霉菌肥防治黄瓜根腐病后,茄病镰刀菌变化趋势一致,初始拷贝数分别为2.61和15.34 copys/μL,7 d内茄病镰刀菌DNA拷贝数增加明显,100×枯草芽孢杆菌和木霉菌DNA拷贝数分别为11.22和20.98 copys/μL,之后茄病镰刀菌数量保持平稳。经过稀释分离法分离2种生防菌和茄病镰刀菌的数量,变化趋势与荧光定量PCR检测趋势相似。因此使生防菌快速定殖是提高其对土传病害防治效果的重要因素之一。

马铃薯干腐病菌和黑痣病菌拮抗芽胞杆菌的筛选及鉴定

从甘肃国营条山农场马铃薯连作试验田的96个小区以五点取样法采集根际土样,稀释平板法在LB培养基上分离得到625株芽胞杆菌菌株。统计分析表明,土壤中总芽胞杆菌的数量正茬>连作2年>连作4年,但连作6年的土壤中总芽胞杆菌的数量又有上升的趋势。以马铃薯干腐病菌接骨木镰刀菌Fusarium sambucinum FSM、茄病镰刀菌F.solani FSI、黑痣病菌立枯丝核菌Rhizoctonia solani RS1和RS2为靶标病原菌,平板对峙培养法筛选有抑菌效果的拮抗芽胞杆菌,发现抑菌带宽度大于2 mm的拮抗芽胞杆菌的数量随连作年限的增加而减少。以形态学特征、生理生化特性为基础,结合gyrB基因序列分析,将筛选出的2株拮抗效果较好且不引起马铃薯块茎腐烂的芽胞杆菌Bacillus spp.D1 0 1和D1 0 2鉴定为萎缩芽胞杆菌B.atrophaeus。

壳聚糖对番茄枯萎病菌的拮抗作用

采用含毒介质法研究了壳聚糖对番茄枯萎病菌菌丝生长和孢子萌发的抑制作用 .结果表明 ,浓度大于 1mg/ m L的壳聚糖能明显抑制菌丝生长 ,各设定浓度均可在一定程度上抑制孢子萌发 ,而且抑制效果随着处理浓度的增大而增强 ;浓度大于 0 .1mg/ m L的壳聚糖处理可诱导初生菌丝发生肿胀、分枝增多且菌体分隔增加及体细胞变短等形态学变化 .壳聚糖的抑菌作用机制与其增加菌丝细胞膜的透性有关 .

根腐病原镰刀菌-苜蓿品种-土壤水分互作对种苗生长的影响

为了探明不同水分、镰刀菌对苜蓿品种的致病性,本研究选择紫花苜蓿品种阿尔冈金(Algonguin)、丰宝(Powerplant)与苜蓿王(Alfaking)接种半裸镰孢(Fusarium semitectum)和腐皮镰孢(F.solani),测定不同水分苜蓿幼苗的病情指数、最终存活数、根重、苗重、根长和苗长,评价其抗病性和两种病菌侵染适宜发病的水分条件以及三者之间的相互关系,结果表明,病菌和水分均对病情指数有显著影响(P<0.05),品种和水分间存在显著的互作关系(P<0.05),品种间病情指数无显著差异,接菌后供试苜蓿品种病情指数均显著高于对照组(P<0.05)。接种腐皮镰孢后显著降低了植株存活数,品种、菌种和水分之间对根重互作效应显著(P<0.05);三者对苗重也有显著影响(P<0.05),且各因素之间也存在显著的互作效应;腐皮镰孢对3个苜蓿品种的苗重均有显著抑制(P<0.05),菌种对根长的影响显著(P<0.05);半裸镰孢对丰宝和苜蓿王品种的根长有较强的抑制作用。品种、菌种和水分对苗长均有显著的影响(P<0.05),而且菌种和水分间存在互作,但品种和菌种、品种和水分、品种、菌种和水分之间互作不显著。接种腐皮镰孢在3个水分梯度条件下对3个品种的苗长均有显著的降低作用。两种镰刀菌在土壤高水分条件下可导致苜蓿发生病害并对苜蓿生产构成危害,腐皮镰孢对苜蓿根部病害的致病性较强。菌种和水分对不同品种的苜蓿病情指数存在显著的影响,而且多因素之间也存在显著的互作效应(P<0.05)。

一株内生拮抗细菌的分离鉴定及其抗菌机理研究

本研究通过平板稀释法、平板对峙法从核桃根部皮层筛选出一株对核桃根腐病有拮抗作用的内生菌株1A,通过形态特征测定、理化特性分析和16SrDNA序列分析鉴定该菌为芽胞杆菌(Bacillus sp.),GenBank序列登录号为KJ865856;其抑菌特性研究结果表明:该拮抗菌对核桃根腐病菌有显著的抑制效果,抑菌率达63.33%;此外,该菌对板栗疫病菌等几种不同的林木病原菌均有较强的抑制作用。抗生机理研究结果表明,该菌株通过分泌蛋白酶和纤维素酶降解真菌细胞壁中蛋白质和纤维素,破坏病菌菌丝,病菌生长受到抑制。该拮抗菌能够显著抑制核桃根腐病菌菌丝生长和孢子萌发,孢子萌发抑制率达74.89%,病菌菌丝表现为菌丝扭曲、断裂,分支增多并缠绕,菌丝颜色加深等。盆栽生防效果研究显示,该拮抗菌对各组核桃根腐病具有显著的防治效果,采用该拮抗菌预处理具有相对较好的生防效果。

稻瘟病生防菌对四种作物土传病害病原菌的抑制作用

利用稻瘟病生防菌（短小芽孢杆菌）对4种主要作物土传病害病原菌进行抑菌作用试验。结果表明，此生防菌对4种作物土传病害病原菌具有不同程度的抑制作用，100倍生防菌发酵稀释液对4种土传病原真菌——辣椒枯萎白绢病菌、节瓜枯萎病菌、香蕉枯萎病病菌、粉蕉根腐病菌生长抑菌率在41．18％以下，显著低于对稻瘟病菌生长的抑制率（69．85％）；生防菌发酵原液对香蕉枯萎病菌的孢子萌发具有较好的抑制作用，抑制率达到86．67％，显著大于其他处理的抑制作用；但不同倍数的生防菌液对节瓜枯萎病菌的抑制差异不显著，抑制率介于76．34％～84．97％之间。

分子标记选择小麦抗白粉病基因Pm4b、Pm13和Pm21聚合体

培育多个抗病基因聚合的小麦品种是提高其抗病广谱性和持久性的有效途径之一。利用小麦抗白粉病基因Pm4、Pm13、Pm2 1的特异PCR标记 ,对含有Pm4b、Pm13、Pm2 1的小麦品系复合杂交F2 代 4 0个植株进行检测 ,从中选择到Pm4b +Pm13+Pm2 13个基因聚合的抗病植株 11个 ,检测、选择到Pm4b +Pm13、Pm4b +Pm2 1、Pm13+Pm2 12个基因聚合的抗病植株 19个 ,为持久、广谱抗病小麦育种奠定了基础。研究还表明 ,3个独立的显性抗病基因在F2 代分离群体中的分离比例基本符合孟德尔独立分配定律 ,在小麦背景下的遗传稳定性 ,与该基因供体和普通小麦的亲缘关系密切相关 ,亲缘关系越远 ,丢失的概率越大。因此 。

枯萎病菌的PCR-RFLP分子检测和早期诊断

采用自行设计的、能对镰刀菌18srDNA进行特异性扩增的1对引物,利用PCR-RFLP对尖孢镰刀菌以不同方式侵染的植株进行跟踪检测。结果表明,应用PCR-RFLP检测程序可检测出处于侵染初期和潜育期的枯萎病菌,说明PCR-RFLP能够快速、灵敏、准确地对枯萎病进行早期诊断。

枝顶孢属真菌的抑菌活性及其代谢产物研究

用组织分离法从采集于广东鼎湖山的腐殖质中分离得到一株枝顶孢属真菌(Acremonium sp.SC0105),其固体发酵物的乙醇提取物对金黄色葡萄球菌(Staphylococcus aureus)有较强的抑制作用。经多种柱层析,从固体发酵物中分离得到5个化合物。通过光谱分析,分别鉴定为姜糖脂B(1)、姜糖脂C(2)、D-甘露醇(3)、酒渣碱(4)、枝顶孢素F(5)。其中酒渣碱是首次从枝顶孢属真菌中分离得到。