寡雄腐霉GAQ1对辣椒疫病的防效及对辣椒的促生作用

为明确自主分离的生防菌株寡雄腐霉Pythium oligandrum GAQ1对辣椒疫病的生防效果及其防御机制,通过平板拮抗和盆栽防效试验测定寡雄腐霉菌株GAQ1对辣椒疫霉Phytophthora capsici菌丝的拮抗作用、对辣椒疫病的防效和对辣椒的促生效果,同时应用实时荧光定量PCR技术检测菌株GAQ1处理后辣椒抗性基因表达的变化。结果表明,寡雄腐霉菌株GAQ1的菌丝可以缠绕并吸附寄生在辣椒疫霉菌丝表面或穿入菌丝体内,使辣椒疫霉菌丝细胞死亡;菌株GAQ1发酵液处理辣椒离体叶片再接种辣椒疫霉后产生的病斑直径较对照组显著减少,离体防效为30.79%;接种菌株GAQ1菌丝球后,辣椒疫病的病情指数较对照组显著降低,盆栽防效达69.16%;经菌株GAQ1处理辣椒后可诱导相关抗性基因PR1、WRKY40、WRKY53、ACCO和GST的相对表达量出现不同程度的升高,说明菌株GAQ1可诱导辣椒植株产生不同程度的防御系统应答。菌株GAQ1对辣椒具有良好的促生效果,处理后第5周其株高、株重及根重分别较对照组提高10.11%、33.23%和24.72%,其叶片中叶绿素a、叶绿素b及类胡萝卜素的含量分...     

寡雄腐霉重寄生作用的研究

 通过室内筛选和盆栽测定证明:寡雄腐霉(Pythium oligandrum)对Rhizoctoniasolanl AG-4,P.ultimum,P.spinosum和P.irregulare有抗生作用,其中以P.oligandr-um的卵孢子处理番茄种子对P.ultimum和P.solani AG-4引起的种腐,猝倒防治效果较好,防效分别为79.4%和63.6%。P.oligandrum对R.solani AG-4主要以菌寄生为主,菌丝能大量分枝侵入,而对P.ultimum主要以抗生为主,分泌出胞壁溶解物质溶解其菌丝。     

寡雄腐霉生防机理及应用研究进展

寡雄腐霉是一种重要的微生物农药,因其对病原菌具有广谱、高效的防治特性及对作物具有促生长和增产等特点被广泛关注。本文从寡雄腐霉对植物病原菌抗性、诱导植物产生诱导系统抗性、促进植物生长等方面对其生防的分子机理和信号转导的研究进展进行了综述,总结了寡雄腐霉在防治植物病害方面的应用,并对研究和应用过程中存在的问题及解决途径提出展望。     

寄生玉米的6种腐霉及其致病性研究

 从国内12省、自治区、直辖市玉米植株上分离出腐霉菌菌株,依据形态特征、培养性状、生长抑制温度,以及产生的有性与无性器官,鉴别出6个腐霉种:棘腐霉(Pythium acanthicum Drechsler),瓜果腐霉(P.aphanidermatum(Edson)Fitzpatrick),强雄腐霉(P. arrhenomanes Drechsler),禾生腐霉(P.graminicola Subramaniam),肿囊腐霉(P.inflatum Matthews),寡雄腐霉(P.oligandrum Drechsler),其中肿囊腐霉出现频率高,分布广泛。回接玉米表明,腐霉能够侵染玉米并引起病害,肿囊腐霉、禾生腐霉、强雄腐霉、棘腐霉是玉米青枯病的重要致病菌;瓜果腐霉能够引起玉米中部茎节腐烂;寡雄腐霉虽寄生玉米,但对玉米成株无致病力。     

木霉菌和腐霉菌对棉铃疫菌的作用研究

用玻璃纸对峙法观察到4种木霉(S-327,BT-12,BT-95,T-594)和寡雄腐霉(P.O.)对棉铃疫霉的营养生长均有一定抑制作用,但没有看到明显的抑菌圈。生物测定试验表明,拮抗菌不仅对棉铃疫霉有直接作用,而且还可以保护棉铃不受疫菌侵染。在5种拮抗菌中,P.O.和BT-12可能是防治棉铃疫病比较有潜力的生防菌。     

寡雄腐霉发酵液对温室番茄生长及灰霉病的防治作用

为了研制高效、无毒的生物农药,采用自主分离的寡雄腐霉生防菌株Pythium oligandrum CQ2010制备发酵液,研究其对温室番茄生长及灰霉病的防治作用。结果表明,寡雄腐霉发酵液(Pythium oligandrum broth,POB)能显著提高番茄叶绿素含量,增强根系活力,增加氮、磷、钾吸收,促进植株生长,使生物量比对照提高65.14%;同时能激活与叶片抗病性相关的过氧化物酶和过氧化氢酶活性,诱导植株产生抗病性反应,提高抗病能力,其效果优于寡雄腐霉卵孢子制剂;而且POB能显著抑制离体灰葡萄孢菌的菌丝生长和孢子萌发;施用POB能使番茄灰霉病发病率下降22.28%~31.05%,病情指数从52.5(对照)下降至22.5~27.5,相对防治效果达47.62%~57.14%,叶片丙二醛含量显著降低,说明POB减轻了灰葡萄孢菌对细胞膜的伤害。表明POB兼具促进番茄生长及防治灰霉病的作用。     

寡雄腐霉发酵液的动物毒性及对辣椒的促生防病效应

为研制高效、无毒、低成本的生物农药以防治辣椒炭疽病,利用自主分离的寡雄腐霉Pythium oligandrum CQ2010菌株制备浓度为1.14‰发酵液,研究其对小鼠的急性毒性、对辣椒幼苗生长的影响及对辣椒炭疽病的防治效果。结果表明,小鼠1日内灌胃寡雄腐霉发酵液60 m L/kg,连续给药14 d后,对小鼠体重增长无显著影响,其外观和行为均无异常,组织器官也未见病理改变。寡雄腐霉发酵液可显著提高辣椒叶片叶绿素含量和根系活力,促进氮、磷、钾吸收,生物量比对照增加42.4%,效果优于寡雄腐霉卵孢子制剂;并能显著提高叶片中过氧化物酶活性,诱导植株抗病性。在辣椒接种炭疽病菌前后,寡雄腐霉发酵液均显著降低了叶片丙二醛的增幅,说明细胞膜受害减轻。施用寡雄腐霉发酵液可使辣椒炭疽病发病率和病情指数显著降低,分别为40.0%~46.7%和13.3~16.7,防治效果达59.9%~68.1%;在模拟自然发病试验中也能显著提高辣椒幼苗生物量,防治效果达55.4%。表明寡雄腐霉发酵液对动物安全无毒,能促进辣椒幼苗生长并防治炭疽病。     

寡雄腐霉发酵液对烤烟生长的影响及对烟草黑胫病的防治作用

为了探索对烟草黑胫病高效、稳定的生防制剂,利用寡雄腐霉工程菌株制备发酵液,采用温室盆栽试验研究该发酵液对烟苗生长的影响和对烟草黑胫病的防治作用。寡雄腐霉发酵液可提高正常烟苗生物量达22.6%,促进烟苗氮、磷、钾吸收;带病烟苗的发病率和病情指数分别降低了61.6%和64.2%,相对防治效果达64.2%;施用发酵液明显降低带病烟苗的细胞膜透性和丙二醛含量,提高烟叶多酚氧化酶、超氧化物歧化酶和苯丙氨酸解氨酶活性。说明寡雄腐霉发酵液可诱导烟株的防御作用,并提高其抗病能力和促生作用。     

紫外光诱导哈茨木霉产生腐霉利抗性菌株的研究

通过紫外光照射结合使用含药培养基培养得到了 3个哈茨木霉腐霉利抗性突变菌株。抗性菌株与亲本菌株相比较 ,其抗药性提高 1 0 0倍以上 ,与异菌脲、菌核净和甲基立枯磷存在交互抗性。抗性菌株在无药培养基上连续转移培养 8次 ,抗性程度未见下降。在适合度上 ,抗性菌株的菌丝生长速率有所下降 ,而在产孢能力、活体抑菌能力上有的抗性菌株要高于亲本菌株。 3个抗性菌株均保持了对灰霉病菌的拮抗能力     

木霉对植物的促生及诱导抗性研究进展

木霉不仅能够直接抑制植物病原菌的生长,还可通过定殖于植物的根部引起植物新陈代谢的改变。本文综述了木霉在植物根部定殖后,通过产生植物生长调节剂、抑制或降解根际有害物质、增加养分利用率以促进植物生长以及通过产生激发子诱导植物形成胞壁沉积物、合成抗菌物质以诱导植物局部或系统抗性的研究进展,以期为进一步扩大木霉在农业生产中的应用提供参考。     

腐霉菌对荧光假单胞菌株LT6代谢物的敏感性变异

为了探明不同靶标病原菌对生防菌荧光假单胞菌株LT6及其代谢物的敏感性差异，在1／5TSA平板上测定了荧光假单胞菌株LT6对69株腐霉菌的拮抗效果，在试验中观察到菌株LT6对不同种腐霉与同种腐霉不同菌株的拮抗效果存在明显差异，抑菌带最宽的为终极腐霉Pythium ultimum菌株，达7．5mm，对未知种腐霉菌株抑菌带宽最小，均小于3．0mm。腐霉菌株对由LT6产生的抗生素吩嗪-1-羧酸（PCA）的敏感性存在变异，被测的54株腐霉，当PCA浓度为7．0μg／mL时，敏感的为28株，高抗的为15株。用不同的培养液培养LT6所得的代谢物，在相同浓度下，对同一腐霉菌菌丝体生长的抑制效果不同，培养液1／5 TSB和NB明显强于KMB。综合所获结果可以看出，靶标病原菌对生防菌敏感性不一致；不同的培养基产生不同的代谢产物，对病原菌的抑制作用有明显差异。     

Induction of transient ethylene and reduction in severity of tomato bacterial wilt by Pythium oligandrum

Pythium oligandrum (PO) is a mycoparasite on a wide range of fungi and suppresses diseases caused by fungal pathogens when colonizing the rhizosphere. PO and its cell wall proteins (CWPs) have elicitor activity that induces defence responses in plants. The potential of a mycelial homogenate of PO to suppress bacterial diseases was studied in roots of tomato ( Lycopersicon esculentum cv. Micro-Tom) inoculated with Ralstonia solanacearum . PO-treated plants showed enhanced resistance to R. solanacearum and reduction in severity of wilt symptoms. As ethylene often acts as one of the signal molecules for induced resistance, its production following treatment of tomato roots with the mycelial homogenate or CWP of PO was measured. The level of ethylene in PO- and CWP-treated plants was transiently elevated six- to 11-fold at 4–8 h after treatment, followed by high expression of three basic ethylene-inducible defence-related genes ( PR-2b , PR-3b and PR-5b ). Analysis of PR-5b gene expression in the leaves of PO- and CWP-treated plants suggested that PR gene expression was induced systemically. The expression of LeERF2 and LeETR4 , which confer an ethylene-dependent signalling pathway, was also significantly accelerated by such treatments. These results indicate that PO has the potential to control bacterial wilt disease and that CWP may play an important role in the induction of resistance to R. solanacearum accompanying the activation of the ethylene-dependent signalling pathway.     

原生质体融合技术改良植病生防木霉菌株

以产孢量大，对苯菌灵有抗性，对潮霉素B敏感的哈茨木霉菌株T9和产孢量少，对潮霉素B有抗性，对苯菌灵敏感的康宁木霉菌株Tk7a为亲本，进行原生质体融合，筛选获得抗最高浓度杀菌剂的融合子。所有融合子的产孢量均比亲本菌株Tk7a大。融合子按照菌落形态的不同分成4组，大部分融合子的几丁质酶、葡聚糖酶和内切葡聚糖酶的活性比亲本菌株高。融合子对棉花猝倒病菌和小麦全蚀病菌的寄生能力基本等同于亲本菌株；融合子对小麦全蚀病的防效比亲本菌株稍好，其刺激麦苗根部生长的能力与亲本菌株相差不大。两个融合子根际竞争能力比亲本菌株T9强，和亲本菌株Tk7a相当。     

邻烯丙基苯酚处理黄瓜种子控制土传腐霉菌和丝核菌侵染危害秧苗的效果

采用ISTA滤纸法检测得出:20%邻烯丙基苯酚悬浮种衣剂按1∶ 30~1∶ 90药种比(质量比)包衣处理"中农8号"和"长春密刺"黄瓜品种的种子,对黄瓜种子安全,种子发芽率分别达到95.75%~97.75%和80.00%~84.25%;PDA平板上检测药剂包衣处理对黄瓜种子带菌消毒效果显示,消毒处理效果达到66.67%~100%。菌丝生长速率法和染色处理结合显微技术实验证实,邻烯丙基苯酚对危害黄瓜秧苗的土传病原菌腐霉病菌Pythium aphanidermatum和立枯丝核菌Rhizoctonia solani具有较强的抑制作用,EC50值分别为27.59和34.16 μg/mL;药剂处理使供试病菌的菌丝发生中空、解体断裂等异常现象。20%邻烯丙基苯酚悬浮种衣剂1∶ 30~1∶ 90包衣处理黄瓜种子后接种供试病菌,对腐霉病菌和立枯丝核菌的防病效果分别达到73.31%~100%和85.16％~100%;随着药剂包衣处理浓度的增大,邻烯丙基苯酚防治两种病原菌侵染危害黄瓜秧苗的效果提高,等效或者优于常规药剂福美双种子处理。     

Biocontrol of potato early blight and suppression of Alternaria grandis sporulation by Clonostachys spp.

Early blight, caused by Alternaria solani and A. grandis, severely affects potato crops around the world, and few management options apart from intensive fungicide use are available. In this work, we tested whether biocontrol treatments with different species of the mycoparasitic fungus Clonostachys could reduce early blight severity on the foliage of pot-grown potatoes of cultivar Ágata, and affect the survival and sporulation of the causal pathogen A. grandis. Twenty isolates from five Clonostachys species were screened in a preliminary biocontrol trial, and nine of them were selected and tested further in two confirmation trials. Treatments with three isolates from three species (C. chloroleuca, C. pseudochroleuca, C. rhizophaga) resulted in a significant reduction in disease severity in both confirmation trials, with control efficacy ranging from 88.7% to 92.9% in the first trial, and from 83.1% to 84.7% in the second trial. All Clonostachys isolates used in the confirmation trials survived on potato leaf tissues for at least 15 days, and resumed growth when subjected to high humidity, resulting in mycoparasitic overgrowth and significant reduction of the pathogen sporulation. Our results provide evidence that different species of Clonostachys, not only the ubiquitous C. rosea, are sources of useful candidates for the biocontrol of potato early blight and possibly other plant diseases caused by fungal pathogens.     

Induced Systemic Resistance (ISR) by Pseudomonas spp. Impairs Pre- and Post-Infection Development of Pythium aphanidermatum on Cucumber Roots

The effect of induced systemic resistance (ISR) by Pseudomonas rhizobacteria on the pre- and post-infection development of Pythium aphanidermatum on cucumber roots was investigated. Cucumber plants (cv. Corona) were grown in vermiculite, roots were split with one side bacterized with Pseudomonas corrugata strain 13 or P. aureofaciens strain 63-28 (bacterized roots) and the other distant side was treated with water (distant, induced roots). For the non-induced control, roots on the bacterized side were treated with buffer instead of the bacterial treatment. Intact, non-split roots were also treated with the bacteria or buffer as a control. Cucumber root tissue from these treatments were harvested and incubated with a zoospore suspension of P. aphanidermatum for three hours. Most of the zoospores in the suspension were stimulated to encyst or germinate. The numbers of germinated zoospores were significantly decreased on distant induced cucumber roots in comparison to non-induced controls. Germination was also reduced on intact bacterized roots, compared to controls. There was less attachment, germ tube production and penetration on roots bacterized or induced by the rhizobacteria compared to non-induced roots. Effects were significantly greater on bacterized roots (roots colonized by bacteria) compared to distant induced roots (roots with the opposite side bacterized). Systemic resistance induced by the two Pseudomonas spp. also reduced pathogen spread on split cucumber roots in planta. Crown infection from induced or bacterized roots was delayed for four to six days in comparison to the non-induced control. Results indicated that Pseudomonas spp. can exert both an indirect influence on P. aphanidermatum zoospore behaviour and infection via induced systemic resistance (ISR) and a local influence via antibiosis or local induced resistance.     

Effect of salinity on pythium damping-off of cucumber and on the tolerance of Pythium aphanidermatum

A study was conducted to investigate the effect of salinity on pythium damping-off and tolerance to salinity among Pythium aphanidermatum isolates from different geographical origins, genetic backgrounds and saline/non-saline soils. Increasing irrigation-water salinity from 0·01 to 5 dS m⁻¹ significantly increased mortality in cucumber seedlings inoculated with P. aphanidermatum and reduced dry weight of non-inoculated seedlings. In vitro tests in culture media amended with different concentrations of NaCl showed that growth of P. aphanidermatum, P. spinosum and P. splendens isolates was stimulated or unaffected at salinity levels stressful for cucumber (electrical conductivity = 5 dS m⁻¹). Significant differences were found in tolerance to salinity among 47 P. aphanidermatum isolates obtained from different greenhouses. The concentration of NaCl required to reduce growth of P. aphanidermatum isolates by 50% varied from 23 to 62 dS m⁻¹, with an average of 46 dS m⁻¹, however, oospore production was more sensitive to salinity and no oospores were produced above 20 dS m⁻¹. Analysis of tolerance of P. aphanidermatum isolates to salinity showed no relationship between tolerance to salinity and geographical origins or genetic data. Isolates of P. aphanidermatum from greenhouses with no salinity problems were as tolerant to salinity as isolates obtained from salinity-affected greenhouses, suggesting lack of evidence for ecological adaptation within P. aphanidermatum in Oman. Increased mortality in cucumber seedlings at higher salinity levels may imply a synergistic interaction between salinity stress and salinity-tolerant Pythium species on cucumber seedlings, resulting in greater seedling losses.     

寡糖诱导植物防卫反应的信号转导

寡糖作为一种生物类激发子,可以诱导植物产生防卫反应,提高植物的抗病性,从而抵御病原物的入侵。本文就寡糖的种类、特点及诱导植物防卫反应信号转导等方面作一评述。     

4株拮抗细菌对棉花黄萎病的防治效果及机制

棉花黄萎病严重制约新疆棉花持续高产和稳产,拮抗细菌在棉花黄萎病生物防治中潜力巨大。本研究旨在明确4株拮抗细菌对棉花黄萎病的防治效果及其机制,为棉花黄萎病的生物防治提供理论依据。本文采用共培养法,研究4株拮抗细菌对黄萎病菌分生孢子浓度、菌丝和孢子形态、膜透性和产毒素能力的影响,采用滤液接种法进行盆栽试验验证其对棉花黄萎病的防治效果,采用愈创木酚法和氮蓝四唑光化还原法测定棉株体内防御酶系的活性。结果表明,4株拮抗细菌能够减少黄萎病菌分生孢子浓度,破坏其细胞形态,导致其细胞膜破裂,降低其毒蛋白浓度。接种4株拮抗细菌,能够诱导棉株体内POD酶和SOD酶等防御酶系的积累,提高了棉花的抗病能力,由菌株SHZ-24、SHT-15、SMT-24和BHZ-29处理的棉花,其对棉花黄萎病的最高防治效果分别为73.82%、80.48%、84.91%和84.18%。4株拮抗细菌通过抑制黄萎病菌的生长和诱导植物产生防御反应来提高对棉花黄萎病的抗性,其对棉花黄萎病具有良好的防治效果。     

Biocontrol and population dynamics of Fusarium spp. on wheat stubble in Argentina

The biocontrol effect of Clonostachys rosea (strains 016 and 1457) on Fusarium graminearum, F. avenaceum, F. verticillioides, F. langsethiae, F. poae, F. sporotrichioides, F. culmorum and Microdochium nivale was evaluated on naturally infected wheat stalks exposed to field conditions for 180 days. Experiments were conducted at two locations in Argentina, Marcos Juarez and Río Cuarto. Antagonists were applied as conidial suspensions at two inoculum levels. Pathogens were quantified by TaqMan real‐time qPCR. During the first year at Marcos Juarez, biocontrol was observed in one antagonist treatment for F. graminearum after 90 days (73% reduction) but after 180 days, the pathogen decreased to undetectable levels. During the second year, biocontrol was observed in three antagonist treatments for F. graminearum and F. avenaceum (68·3% and 98·9% DNA reduction, respectively, after 90 days). Fusarium verticillioides was not controlled at Marcos Juarez. At Río Cuarto, biocontrol effects were observed in several treatments at different intervals, with a mean DNA reduction of 88·7% for F. graminearum and F. avenaceum, and 100% reduction for F. verticillioides in two treatments after 180 days. Populations of F. avenaceum and F. verticillioides were stable; meanwhile, F. graminearum population levels varied during the first 90 days, and low levels were observed after 180 days. The other pathogens were not detected. The study showed that wheat stalks were important reservoirs for F. avenaceum and F. verticillioides populations but less favourable for F. graminearum survival. Clonostachys rosea (strain 1457) showed potential to reduce the Fusarium spp. on wheat stalks.