Increased Incidence of Erwinia Soft-rot on Calla Lilies in the Presence of Phosphorous

Erwinia soft-rot is an important disease of many ornamental potted crops and is one of the most limiting factors in greenhouse calla lily (Zantedeschia spp.) production. Experiments were conducted to test the effect of phosphorous added to soil-less mixes or to nutrient solutions used for irrigation on soft-rot caused by Erwinia carotovora subsp. carotovora (Ecc). Soft-rot incidence increased to 51% when soil-less mix was amended with superphosphate in comparison to regular soil-less mix (no superphosphate added) (31%). In contrast, addition of phosphorous in the nutrient solution met the phosphorous needs of the plant without enhancing soft-rot. Plant height, fresh mass, and number of flowers per plant were greater in calla lilies irrigated with nutrient solution containing phosphorous than no phosphorous treatments. Similar results were obtained in tests conducted in a commercial greenhouse with larger sample size. No statistical differences were found between tubers sprayed with water (control) or with a 0.5 mM solution of KH₂PO₄ in laboratory experiments to determine the effect of phosphorous on tuber root development. In other experiments, tubers were sprayed with either water, a bacterial cell suspension 1 × 10² cfu ml^–1, a solution of 0.5 mM KH₂PO₄, or a suspension of bacteria in KH₂PO₄. The results from these tests showed a significant increase of soft-rot development in tubers treated with the suspension of Ecc prepared in a solution of KH₂PO₄ relative to other treatments. Further laboratory tests indicated that enzymatic activity (polygalacturonase and pectate lyase) of Ecc increased when grown in the presence of phosphorous. These experiments suggest that increased soft-rot in the presence of phosphorous is due to increased virulence of Ecc.     

Characterization of an antibacterial substance produced by <Emphasis Type="Italic">Erwinia carotovora</Emphasis> subsp. <Emphasis Type="Italic">carotovora</Emphasis> Ecc 32

A total of 88 strains of Erwinia carotovora subsp. carotovora (Ecc) isolated from various host plants in several geographic regions were screened for production of antibacterial substances using the same strains as indicators. Of the 88 strains, 72 produced antibacterial substances. One of these 72 strains, a Brazilian strain Ecc 32, produced an antibacterial substance active against all tested Ecc strains on TSA medium. The antibacterial spectrum of the compound from Ecc 32 strain was limited to closely related strains of soft-rot Erwinia species. Such a narrow spectrum of activity is typical of bacteriocins. The compound produced by Ecc 32 strain, however, was resistant to some enzymes and detergents. Moreover, the compound was heat-stable and active over a wide pH range. The physical characteristics of the compound were not in agreement with those of bacteriocin or carotovoricin.     

十字花科根肿病生防菌副地衣芽胞杆菌ZF480的鉴定及其防效

为有效防控十字花科蔬菜生产中的毁灭性病害——根肿病,以尖孢镰孢菌Fusarium oxysporum和辣椒疫霉Phytophthora capsici为指示菌,利用平板对峙法对分离自土壤的1 198株细菌进行筛选,进通过盆栽试验筛选对十字花科根肿病具有良好防效的生防菌株,并基于形态学特征、生理生化特性及16S rDNA、gyrB和atpD多基因系统发育分析对生防菌株进行鉴定,同时测定其抑菌谱和田间防效。结果表明,通过对峙培养筛选获得115株有抑菌效果的生防菌株,采用盆栽试验获得1株对根肿病菌具有拮抗效果的菌株,命名为ZF480。结合形态学特征和系统发育分析结果将该菌株鉴定为副地衣芽胞杆菌Bacillus paralicheniformis。菌株ZF480对野油菜黄单胞野油菜变种Xanthomonas campestris pv. campestris等10种病原菌具有拮抗效果,抑菌谱较宽,对白菜根肿病的盆栽防效和田间防效分别达到72.82%和64.08%。表明副地衣芽胞杆菌菌株ZF480对十字花科根肿病具有显著的防效,可作为生防菌进一步开发利用。     

Moss-Erwinia pathosystem reveals possible similarities in pathogenesis and pathogen defense in vascular and nonvascular plants

Vascular plants have various inducible resistance mechanisms as defense against pathogens. Mosses, small nonvascular plants (subkingdom Bryophyta), have been little studied in regard to their pathogens or modes of defense. Data here show that Erwinia carotovora, a bacterial plant pathogen that causes softrot in many dicotyledonous plants, can also cause soft rot symptoms in the moss Physcomitrella patens. Infection of moss by E. carotovora required pathogenicity factors similar to those required to infect vascular plants and, again as in vascular plants, salicylic acid (SA) induced moss to inhibit tissue maceration by Erwinia. These data reveal that SA-dependent defense pathways may have evolved before differentiation of vascular and nonvascular plants.     

Characterization and differentiation of <Emphasis Type="Italic">Erwinia carotovora</Emphasis> strains from mulberry trees

We recently reported that two diverse types (types 1 and 2) were identified among strains of Erwinia carotovora from mulberry trees. Type 1 strains were similar to E. carotovora subsp. carotovora (Ecc), whereas type 2 strains were distinct from Ecc and other E. carotovora strains. In this study, seven more mulberry strains of type 2 and reference strains were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and randomly amplified of polymorphic DNA (RAPD). On the basis of SDS-PAGE profiles of whole-cell proteins, type 2 strains had high similarity with one another. In addition, they had an unique peptide band with a molecular mass of approximately 28kDa. RAPD analysis showed that they were also effectively differentiated by a strong, specific RAPD fragment for type 2 strains. Based on these two approaches, we have confirmed that the present type 2 strains from mulberry can be discriminated clearly from other soft rot Erwinia species.     

海洋生境贝莱斯芽孢杆菌TCS001的鉴定及抑真菌活性

为研究海洋生境芽孢杆菌TCS001的分类地位和抑菌活性,通过形态和生理生化特征观察,并结合gyrA序列同源性分析对菌株进行了鉴定;通过平板对峙培养法测定了菌株TCS001对多种植物病原真菌的抑菌谱;采用菌丝生长速率法和凹玻片法,测定了不同浓度TCS001菌株发酵滤液对靶标菌黄瓜灰霉病菌Botrytis cinerea菌丝生长和孢子萌发的影响。结果显示:该菌株为贝莱斯芽孢杆菌Bacillus velezensis,其对6种供试病原菌均有一定的抑制效果,其中对黄瓜灰霉病菌的抑制率最高,达87.66%。不同稀释倍数下,TCS001发酵滤液对黄瓜灰霉病菌均有一定的抑制作用,其中,稀释5倍时对菌丝生长和孢子萌发的抑制率最高,分别为96.24%和98.05%,稀释20倍时抑制率也均达90%以上。形态学观察发现,TCS001发酵滤液可导致黄瓜灰霉病菌孢子萌发芽管中间或顶端膨大畸形。研究表明,海洋生境贝莱斯芽孢杆菌TCS001极具开发为微生物农药的潜能。     

Flagella-mediated motility is required for biofilm formation by Erwinia carotovora subsp. carotovora

To elucidate the role of flagella in biofilm formation by Erwinia carotovora subsp. carotovora EC1, we used a nonflagellate, nonmotile mutant (ΔfliC) and a flagellate, nonmotile mutant (ΔmotA). A biofilm-inducing medium, which contains the yeast peptone (YP) medium plus the salts of M-63 minimal medium, supported biofilm formation to a greater extent than either the YP or Luria Bertani (LB) medium alone. We demonstrated that both the ΔfliC and ΔmotA mutants greatly reduced their ability to form a biofilm on the surface of the wells of polyvinyl chloride (PVC) microtiter plates. The inability of both mutants to form biofilm on the PVC surface was further confirmed with phase-contrast microscopy. Both aflagellate (ΔfliC) and flagellate (ΔmotA) nonmotile mutants were equally defective in attachment to the PVC surface. The treatment of bacteria with the protonophore carbonyl cyanide m-chlorophenylhydrazone (CCCP), which inhibits the motility of this organism, reduced greatly the biofilm formation. Based on these results, flagella-mediated motility may play an important role in biofilm formation of E. carotovora subsp. carotovora EC1.     

特基拉芽孢杆菌JN-369的分离鉴定及其抑菌物质分析

为筛选可用于防治由稻瘟病菌所致水稻稻瘟病的生防菌株资源,采用稀释涂布平板法,从感病水稻品种湘早籼24号的健康植株茎叶中分离获得了1株拮抗细菌JN-369,结合形态学观察、生理生化鉴定及16S rDNA序列分析对其菌种进行了鉴定,采用平板对峙法研究了JN-369菌株的抑菌谱,并初步测定了JN-369中挥发性有机物 (VOCs)、蛋白类粗提物及脂肽类粗提物对稻瘟病菌的抑制作用。结果表明:JN-369菌株为特基拉芽孢杆菌Bacillus tequilensis,其对稻瘟病菌具有显著抑制作用,对菌丝生长的抑制率达80.46% ± 0.83%;同时该菌株对供试的辣椒胶孢炭疽菌、烟草赤星病菌及黄瓜疫病菌等植物病原真菌和卵菌均有抑制作用;但对供试病原细菌则均无抑制作用;1 × 108 cfu/mL 的 JN-369菌悬液产生的挥发性有机物对稻瘟病菌的抑制效果最强,抑制率达72.92% ± 3.01%,1.454 mg/mL的蛋白类提取物和1.026 mg/mL 的脂肽类提取物对稻瘟病菌的抑制率分别为24.68% ± 0.80%和14.34% ± 1.08%。研究表明,菌株JN-369具有一定的开发应用潜力。     

木霉分生孢子和厚垣孢子对黄瓜幼苗生理特性及枯萎病防效的影响

采用前期筛选出的对黄瓜枯萎病菌有较好拮抗作用的3株木霉菌,即哈茨木霉Trichoderma harzianum 809、拟康氏木霉Trichoderma pseudokoningii 886和棘孢木霉Trichoderma asperellum 525,利用盆栽试验,测定了木霉菌孢子不同类型施用对黄瓜幼苗生长、生理特性及枯萎病防效的影响。结果表明:哈茨木霉809、拟康氏木霉886、棘孢木霉525分生孢子和厚垣孢子对黄瓜枯萎病的盆栽防效均在66.81%以上,且以拟康氏木霉886厚垣孢子防效最高,达到81.46%。当黄瓜幼苗长至三叶一心时,3株木霉菌分生孢子和厚垣孢子处理的黄瓜幼苗株高、茎粗、根体积、叶面积、全株鲜质量均比CK显著上升,以厚垣孢子886的促进效果最显著,其株高、茎粗、根体积、叶面积、全株鲜质量分别较CK增加了85.33%、108.43%、235.29%、144.38%、272.32%。同时,3株木霉菌分生孢子和厚垣孢子处理的黄瓜幼苗叶绿素含量、根系活力、叶片可溶性糖含量、叶片可溶性蛋白含量也均比CK显著上升,厚垣孢子886的促进效果仍最显著,其叶绿素含量、根系活力、叶片可溶性糖含量、叶片可溶性蛋白含量分别较CK增加了97.47%、86.05%、172.49%、201.91%。     

斯氏线虫-致病杆菌共生体侵染昆虫的生物标志物

昆虫病原线虫是一类专性侵染和寄生昆虫的病原线虫,是非常重要的生防资源。斯氏线虫属Steinernema和致病杆菌属Xenorhabdus通过形成共生体在侵染昆虫过程中共同完成生活史,其中致病杆菌释放效应物质引发昆虫败血症是其重要机制。该文对斯氏线虫-致病杆菌共生体的侵染策略和在侵染过程中产生的免疫调节因子、释放的毒素蛋白和活性代谢物进行概述,其中,苯乙酰胺作为免疫抑制因子促进自身定殖,脂多糖作为内毒素引起寄主血细胞裂解,Tc毒素蛋白作为外毒素导致寄主中肠上皮细胞溶解,活性代谢产物如xenematides、fabclavine和PAX肽等具有抑菌、诱导细胞凋亡等活性。而斯氏线虫本身也能够产生表皮/分泌蛋白来抑制寄主免疫,与共生菌协同致死寄主。因此,通过对斯氏线虫-致病杆菌共生体产生的致病物质进行汇总分析,为研究昆虫病原线虫致病机制提供理论参考,同时也为新型绿色杀虫剂的开发和应用提供证据支持。     

棉花黄萎病拮抗细菌H14的筛选鉴定及其拮抗机理分析

为筛选对大丽轮枝菌Verticillium dahliae Kleb.具有拮抗活性的根际细菌,以大丽轮枝菌为靶菌,分离获得棉花根际细菌,利用平板对峙法和琼脂扩散法筛选具有较高拮抗活性的菌株,采用室内盆栽法测定筛选所得菌株对棉花黄萎病的防效,并通过形态特征、生理生化特性和16S r DNA基因序列分析确立其分类地位,采用底物降解法和抗菌肽基因克隆法检测其产生水解酶和抗菌肽的能力。结果显示,试验共分离获得182株棉花根际细菌,筛选得到3株对大丽轮枝菌抑菌率大于50.00%且抑菌圈直径大于15 mm的菌株,其中菌株H14的抑菌率和抑菌圈直径最大,分别为54.25%和18.10 mm。该菌能在0～9%NaCl和pH 4.5～9.0的NB培养基上生长;经形态特征观察、生理生化试验及16S rDNA基因序列分析,最终将其鉴定为莫哈韦芽胞杆菌Bacillus mojavensis;菌株H14对大丽轮枝菌孢子萌发的抑制率和对棉花黄萎病的盆栽防效分别为89.55%和74.57%;菌株H14能合成蛋白酶,含有srfAA、fenD、bacA脂肽类抗生素合成基因。表明莫哈韦芽胞杆菌菌株H14能够合成蛋白酶和脂肽类拮抗物质,具有良好的抑菌和防病能力。     

Defense strategies of pea embryo axes with different levels of sucrose to Fusarium oxysporum and Ascochyta pisi

The aim of this study was to compare the defense responses of embryo axes of Pisum sativum L. cv. Kwestor with different sucrose levels to pathogenic fungi, i.e. systemic acting Fusarium oxysporum f. sp. pisi and locally acting Ascochyta pisi. Embryo axes were cultured on Heller medium for 96 h. Four variants were compared: these included inoculated embryo axes cultured with or without 60 mM sucrose (+Si and −Si) and non-inoculated embryo axes cultured with or without 60 mM sucrose (+Sn and −Sn). After inoculation of the pea embryo axes with pathogenic fungi a generally higher concentration of free radicals was detected by electron paramagnetic resonance (EPR), in comparison to non-inoculated embryo axes. The inoculation with F. oxysporum caused stronger generation of free radicals in −Si than in +Si embryo axes. A different response was observed after inoculation with A. pisi; starting from 48 h, the concentration of free radicals in +Si axes was found to be 1.5 times higher than in −Si embryo axes. The values of spectroscopic splitting coefficients for these radicals suggest that they are semiquinone radicals. The EPR method also revealed Mn²⁺ ion accumulation after 24 h of culture. Over time, high levels of these ions were recorded in +Si embryo axes inoculated with F. oxysporum, while in +Si embryo axes inoculated with A. pisi they decreased. Up to 48 h after inoculation with the pathogenic fungi, Mn²⁺ ion levels were higher in +Si embryo axes than in +Sn axes. The activity of superoxide dismutase (SOD, EC 1.15.1.1) increased in +Si embryo axes up to 72 h after inoculation with pathogenic fungi; however, it was generally lower than in +Sn axes. Catalase activity (CAT, EC 1.11.1.6) increased up to 72 h after inoculation with F. oxysporum and the values were higher than in the non-inoculated tissue. Especially high activity of this enzyme was noted in −Si embryo axes after inoculation with either F. oxysporum or A. pisi. Peroxidase activity (POX, EC 1.11.1.7) towards pyrogallol in embryo axes increased during culture; however, it was lower or similar to that in non-inoculated embryo axes. SOD, CAT and POX zymograms showed that the synthesis of new isoforms was induced after inoculation with pathogenic fungi. Peroxidase isozymes detected by the reaction with diaminobenzidine in native PAGE were intensely stained in +Si embryo axes after inoculation with pathogenic fungi. Respiratory activity of the inoculated tissues was considerably higher than in non-inoculated tissues. The respiration rate was generally much higher in +Si than in −Si embryo axes. Growth of −Si embryo axes was more significantly retarded as a consequence of inoculation than that of +Si embryo axes.These results indicate that, depending on the manner of influence of a pathogenic fungus, both similar and differing defensive strategies may be initiated and a raised sugar levels in pea tissues limit the development of F. oxysporum and A. pisi.     

一株对桃蚜有高致病性球孢白僵菌的分离、筛选与鉴定

为筛选出对桃蚜Myzus persicae具有高致病性的生防真菌,以从陕西省秦岭原始森林采集到的鳞翅目僵虫虫体中分离获得的5株真菌为研究对象,测定其对桃蚜的致病性并筛选出高致病性菌株表现最佳杀蚜活性时的孢子悬浮液浓度,同时结合形态学及18S rDNA和ITS-rDNA序列分析对致病性最高的病原菌进行鉴定。结果显示,从5株菌株中初步筛选出1株高致病性菌株BQ-63,处理7 d后桃蚜的死亡率为80.33%,校正死亡率为81.58%,僵虫率也达到最高,为80.78%;当菌株BQ-63的孢子浓度为108个/mL时,对桃蚜的致病性达到最高,处理7 d后死亡率为89.53%,校正死亡率为90.10%,僵虫率为89.84%;通过形态学和分子生物学鉴定,确定菌株BQ-63为球孢白僵菌Beauveria bassiana。表明菌株BQ-63对桃蚜具有高致病性,可作为生防真菌进行进一步的研究。     

木霉菌颗粒剂对玉米茎腐病防治的应用

为明确棘孢木霉菌Trichoderma asperellum不同颗粒剂配方对玉米茎腐病的防效,通过人工接种盆栽试验比较分析其防效,同时进行多基点田间试验以筛选对茎腐病防效明显并具有增产效应的木霉菌生防颗粒剂。结果表明:盆栽试验中,2号木霉菌颗粒剂对于苗期玉米茎腐病的防效最高,为65.57%,3号木霉菌颗粒剂对于成株期玉米茎腐病的防效最好,为40.00%;1~4号4种木霉菌颗粒剂均能提高玉米的出苗率、株高、鲜重、干重和根长,表明木霉菌颗粒剂对玉米生长具有促进作用。在全国10余个综合试验站对1号木霉菌颗粒剂的田间防效试验结果显示,对玉米茎腐病的防效较好,均在50%以上,最高可达80%,玉米增产效果在8%以上。表明木霉菌颗粒剂具有良好的生物防治效果及增产作用。     

Use of some plant extracts to control Biomphalaria alexandrina snails with emphasis on some biological effects

The present work was carried out to evaluate, the molluscicidal activity of cold water, boiled water, methanol, ethanol, acetone and chloroform extracts of some plant species against Biomphalaria alexandrina snails. Preliminary screening tests on 10 plant species showed that the highest molluscicidal potency was recorded for Euphorbia splendens, Atriplex stylosa and Guayacum officinalis. Exposure of B. alexandrina snails to plant’s methanol extracts led to a significant reduction in their survival and growth rates. In addition, newly hatched snails were susceptible to plant’s methanol extracts than elder ones. LC₂₅ of methanol extract from these plants caused a considerable reduction in the infectivity of Schistosoma mansoni miracidia to the snails. It caused a reduction in number of cercariae per snail during the patent period and in the period of cercarial shedding. The results, also, revealed that the glucose concentration in treated snails was increased in haemolymph, while soft tissue glycogen decreased. The activities of glycogen phosphorylase, succinate dehydrogenase (SDH), glucose-6-phosphatase and acetylcholinesterase (AchE) in homogenate of snail’s tissues were reduced (P < 0.001) in response to treatment with plants methanol extract, while glucose-6-phosphate dehydrogenase (G-6-PD) activity increased (P < 0.001). It was concluded that the application of LC₂₅ of methanol extracts of E. splendens, A. stylosa and G. officinalis may be helpful in snail control as it interferes with the snails’ biology and physiology.     

蚜虫病原细菌的分离鉴定及其对褐飞虱的杀虫活性初探

为寻找有效防治褐飞虱的病原微生物,采用平板划线法和稀释涂布法,从感病死亡的蔷薇蚜虫体中分离筛选出一株对褐飞虱具有较高致病性的菌株JSA1;通过形态学特征、生理生化特性及16S r DNA序列分析,对菌株JSA1进行了初步鉴定;采用盆栽法和稻苗浸渍法,对其杀虫活性进行了测定。鉴定结果显示菌株JSA1为铜绿假单胞菌Pseudomonas aeruginosa。杀虫活性测定结果表明:菌株JSA1对褐飞虱具有杀虫作用,且致死率随菌液浓度的升高和作用时间的延长而提高,但致死率低于40%;菌株JSA1发酵液的上清液和沉淀破碎液对褐飞虱3龄若虫均具有致病力,72 h后的校正死亡率分别为30.72%和82.53%,表明铜绿假单胞菌JSA1在褐飞虱的生物防治方面具有一定的应用前景。     

淡紫拟青霉QLP12对感染灰霉病后番茄植株的促生作用及抗病相关酶活性变化

为探究淡紫拟青霉Paecilomyces lilacinus菌株QLP12对番茄植株促生作用及番茄灰霉病的防治效果,采用平板对峙法测定菌株QLP12对番茄灰霉病菌即灰葡萄孢Botrytis cinerea的抑制作用,用离体叶片、果实、植株接种法研究其对番茄灰霉病的防效,并对盆栽番茄的促生效果、防御酶活性、丙二醛(MDA)和脯氨酸含进行测定。结果表明,菌株QLP12发酵滤液对灰葡萄孢菌丝生长的抑制率为58.97%,对分生孢子萌发的抑制率为89.28%;盆栽试验结果显示,菌株QLP12发酵液对番茄灰霉病的防效为50.20%;在灰葡萄孢胁迫下,菌株QLP12处理可诱导植株叶片组织防御酶活性和脯氨酸含量升高,丙二醛含量降低,同时促进番茄幼苗生长,株高、茎粗、干鲜比和叶绿素含量与对照相比均有显著提高。表明生防菌QLP12可通过诱导植株防御酶活性的提高,有效抑制番茄灰霉病发生,并促进番茄幼苗生长。     

番茄枯萎病菌和青枯病菌拮抗细菌的评价

为筛选出对番茄枯萎病和青枯病有较好防效的生防菌,采用平板对峙法,以番茄枯萎病菌Fusarium oxysporum和番茄青枯病菌Ralstonia solanacearum为靶标菌,从江苏沭阳、宿迁、溧水及内蒙古海拉尔分离到的2 062株细菌菌株中筛选拮抗菌株,并采用平板对峙法、拮抗菌液灌根法、分子生物学方法进行拮抗物质检测、盆栽试验及种属鉴定。结果表明:从2 062株细菌中共筛选到21株对番茄枯萎病和青枯病具有很强拮抗作用的菌株,均能分泌蛋白酶,具有解磷作用;不能分泌几丁质酶和纤维素酶,仅4株细菌能分泌嗜铁素。拮抗细菌SY290对番茄枯萎病和番茄青枯病防效最高,分别达到74.2%和75.0%,SQ728和LS536次之,但防效均大于60%。结合各菌株形态特征、16S r DNA与gyr-B序列分析结果,菌株SY177、SY290和SQ728鉴定为解淀粉芽胞杆菌Bacillus amyloliquefaciens,菌株LS536为枯草芽胞杆菌B.subtilis。     

Isolation of differentially expressed genes during interactions between tomato cells and a protective or a non-protective strain of Fusarium oxysporum

Pathogenic isolates of Fusarium oxysporum applied on a non-host plant species, as soil-borne non-pathogenic isolates, are able to protect this plant against pathogenic strains inducing wilts. Several modes of action contribute to the biocontrol activity of these protective strains; however the genetic basis of the biocontrol mechanisms is far from being understood. The aim of this study was to identify genes involved in biocontrol activity of F. oxysporum using an original model made of Fom24, a strain protective on tomato and its mutant rev157 which has lost its protective capacity. A Rapid Subtractive Hybridization (RaSH) approach was chosen to identify genes up-regulated in the protective or in the non-protective interaction when germinated conidia of either Fom24 or rev157 are confronted to tomato cell cultures. A total of 86 up-regulated sequences were generated, 42 and 44 from the protective and the non-protective interaction respectively. Homology searches led to identification of both plant and fungal genes that were grouped according to their putative functions. Among plant genes, those involved in plant response to stresses were the most abundant. Expression profiles of genes homolog to a basic endochitinase, a ferredoxine-NADP(H) reductase (FNR), an ATP synthase and the RPM1-interacting protein 4 (RIN4) were confirmed by Northern blotting. A large proportion of fungal sequences were encoding genes of unknown function; among other, those involved in response to oxidative stress and a gene putatively encoding an enolase are the most promising to further study their potential role in the protective interaction between F. oxysporum and tomato.     

生防菌株ZA1的GFP基因标记及其功能稳定性测定

为明确生防莫海威芽胞杆菌Bacillus mojavensis菌株ZA1在作物体内定殖与作物和病原物的作用机理,运用电击转化法将绿色荧光蛋白表达载体pGFP78导入菌株ZA1体内,并采用细菌培养法、继代培养法和平板抑制法检测其功能稳定性。结果表明,质粒pGFP78成功导入菌株ZA1,标记后菌株ZA1-gfp经形态学和分子生物学验证均证明菌株ZA1已成功标记,标记菌株ZA1-gfp与菌株ZA1形态一致,在荧光显微镜下呈现明亮的绿色,并且能够提取导入质粒pGFP78,扩展出质粒所携带的启动子78序列。功能稳定性鉴定结果表明,标记菌株ZA1-gfp遗传稳定性达100%;与菌株ZA1的生长速率差异不显著,且不影响菌株的运动性;菌株ZA1对马铃薯枯萎病菌、马铃薯炭疽病菌和马铃薯坏疽病菌的抑制率分别为64.50%、68.35%和66.55%,标记菌株ZA1-gfp对3种病原真菌的抑制率分别为67.46%、76.56%和66.75%,表明外源质粒的导入,不影响菌株的抑菌能力。