番茄灰霉病菌拮抗基因工程菌株Fh-chib培养条件的研究

[目的]筛选番茄灰霉病菌拮抗基因工程菌株Fh-chib的最佳培养条件。[方法]在单因素试验的基础上,采用正交试验设计对番茄灰霉病菌拮抗菌株Fh-chib的最佳培养条件(碳源、氮源和无机盐)进行了研究。[结果]工程菌株Fh-chib的最佳培养基配方为0.50%葡萄糖、1.00%蔗糖、1.00%酵母膏、1.50%麸皮、1.00%黄豆饼粉、0.03%MgSO4和0.40%CaCl2。各个因素对发酵液抑菌活性影响的主次顺序为:葡萄糖>蔗糖>CaCl2>黄豆饼粉>酵母膏>麸皮>MgSO4。[结论]为该菌株的工业化生产奠定了基础。     

越橘灰霉病病原菌鉴定及其生物学特性研究

对越橘灰霉病进行了病原鉴定、生物学特性及其室内药剂筛选研究.形态特征鉴定与rDNA-ITS序列分析结果表明:Bf.jlnd09菌株鉴定为灰葡萄孢菌(Botrytis cinerea,GenBank登录号为GU395993).生物学特性测定结果显示:病原菌菌丝生长和产孢最适培养基为PDA,菌丝生长最适碳氮源分别为甘露糖和蛋白胨,而产孢最适碳氮源为葡萄糖和蛋白胨,4～35℃条件下均可生长并产孢(最适20℃),病菌在pH为4.0～10.0条件下均能生长并产孢(最适5.0);孢子萌发最适pH为6.0,最低相对湿度为80％,光暗交替有利于孢子的萌发;分生孢子的致死温度为37℃.室内毒力测定结果表明:60％金万霉灵WP、70％甲基托布津WP和53％金雷多米尔GR 3种杀菌剂对越橘灰霉病菌的抑菌效果显著高于其他药剂.     

不同氮磷钾配比对仙客来生长的影响

对仙客来无土栽培营养液中N、P、K元素采用三因素二次D-饱和最优设计,探讨不同浓度氮磷钾对仙客来生长发育的影响,对仙客来的根长、根体积、球茎直径、花朵数、花瓣长、可溶性糖含量、MDA含量、蛋白质含量、叶绿素含量进行分析。结果表明:经方差分析和主成分分析得出,无土栽培仙客来氮、磷、钾最优组合方案为:86.6~263.4、60~100和250~400 mg/L,即氮:磷:钾为3∶1∶6.5。     

An inhibitor of polyamine biosynthesis — Difluoromethylornithine — and the polyamine spermidine for the control of gray mold(Botrytis Cinerea)

Difluoromethylornithine (DFMO) — an inhibitor of polyamine biosynthesis, and the polyamine spermidine (Spd) reduced gray mold of tomato, pepper, eggplant, bean andSenecio sp. leaves, and of rose petals by 37–88% when applied at 0.1–1.0 mM each. Higher doses did not result in better control. The disease was also reduced significantly on tomato fruits by 1.0 inM DFMO and by 0.1–1.0 mM Spd, and on cucumber fruits by 0.1–1.0 mM of both compounds, but not on grape berries. The combination of 0.2 mM DFMO with 1.0 mM Spd controlled gray mold ofSenecio sp. and tomato leaves additively better than either treatment alone, whereas this effect was not observed in leaves of lettuce and pepper. Ethylene production was reduced significantly by Spd applied to leaves of tomato and pepper, but not by DFMO. Linear growth and germination of the fungus were affected by lower concentrations of DFMO (ED50 0.12–0.97 and 1.4, respectively) as compared with Spd. Spermidine and DFMO controlled white mold(Sclerotinia sclerotiorum) as effectively as did the fungicide benomyl. Contribution from the Agricultural Research Organization. No. 3195-E, 1991 series.     

草莓灰霉病菌的培养、毒素提取及生物测定

 草莓灰霉病菌(Botrytis cinerea Persoon)在25℃、pH3~4的Peberdy培养基中生长良好,并可产生引起草莓愈伤组织细胞死亡的毒素。产生毒素的高峰期为静置培养20天。灰霉菌发酵液中的毒素可用氯仿进行提取,经提取后的毒素液在269nm处有吸收峰。本研究建立了利用草莓愈伤组织细胞的荧光活性测定毒力的方法。     

Studies on red light-induced resistance of broad bean to Botrytis cinerea: I. Possible production of suppressor and elicitor by germinating spores of pathogen

When detached broad bean leaves were preinoculated with virulent strain B304 of Botrytis cinerea 24 h before a challenge inoculation with strain B304, lesion formation by B304 was significantly inhibited in red light but not in the dark. In leaves that were preinoculated with avirulent strain 021 and then challenged by B304, however, lesion formation was not inhibited even under red light. Such differences in lesion formation after the challenge inoculation with an avirulent strain were also observed with lesions caused by Alternaria alternata, a nonpathogen of broad bean and by avirulent strain 021R in the presence of germination fluid from spores of strains B304 and 021R. These results suggest the possibility that virulent B. cinerea produced a suppressor involved in induced susceptibility and an elicitor involved in resistance induced by red light during spore germination.     

Local and systemic resistance to fungal pathogens triggered by an AVR9-mediated hypersensitive response in tomato and oilseed rape carrying the Cf-9 resistance gene

Tomato and transgenic oilseed rape plants expressing the Cf-9 resistance gene develop a hypersensitive response (HR) after injection of the corresponding Avr9 gene product. It was investigated whether induction of a HR conferred resistance to different fungal pathogens in tomato and oilseed rape. Induction of an AVR9 mediated HR at the pathogen infection site delayed the development of the biotrophs Oidium lycopersicum in tomato and Erysiphe polygoni in oilseed rape, but enhanced the development of the necrotrophs Botrytis cinerea and Alternaria solani in tomato and Sclerotinia sclerotiorum in oilseed rape. Interestingly, delayed fungal disease development was observed in plant tissues surrounding the HR lesion regardless of whether a necrotrophic or biotrophic pathogen was used. In tomato, AVR9 injection induced systemic expression of PR1, PR2 and PR3 defence genes but did not induce systemic resistance to O. lycopersicum, B. cinerea or A. solani. In oilseed rape, AVR9 injection temporarily induced systemic resistance to Leptosphaeria maculans and E. polygoni, but did not induce detectable systemic expression of PR1, PR2 or Cxc750. These results give new insights into the potential uses of an induced HR to engineer disease resistance.     

Biological control of botrytis leaf blight of onion byGliocladium roseum applied as sprays and with fabric applicators

Gliocladium roseum (5×10⁶ conidia ml^–1) and chlorothalonil (Bravo 500) were compared in two field tests for effectiveness in suppressing leaf blight caused byBotrytis squamosa in cooking onions. The biological control agent and fungicide were applied with sprayers and with specially designed fabric applicators that were mounted interchangeably on an aluminium carriage with bicycle wheels. In the applicators, inoculum and fungicide gravitated from a reservoir down curtains of denim strips and onto portions of onion leaves contacted by the strips. Initial applications were timed using a leaf blight forecasting system (BOTCAST) and three or four subsequent applications were made at weekly intervals. When compared to water checks,G. roseum applied as sprays or by applicators reduced density of leaf spots by about 50–58% during middle and late stages of epidemics. The antagonist was about half as effective as chlorothalonil in suppressing density of spots, regardless of application method. The applicators delivered inoculum in similar densities to proximal and distal halves of leaves, and used at least 35% less volume of inoculum compared to the sprayers. Density of inoculum on leaves treated by applicators was about the same as in spray-treated leaves when canopies were sparse, but less when canopies were dense. It is concluded thatG. roseum has good potential for controlling leaf blight sufficiently to avoid economic yield losses.     

Mode of action of the anilino-pyrimidine fungicide pyrimethanil. 2. Effects on enzyme secretion in Botrytis cinerea

The effect of pyrimethanil on the levels of cell wall degrading enzymes secreted by Botrytis cinerea Pers. was investigated in diseased plant tissues and in liquid B. cinerea cultures. Total proteinase activity isolated from infected carrot slices which were treated with 5.0 μM pyrimethanil was decreased by 76%, 3 d after inoculation. Polygalacturonase, cellulase, proteinase and laccase activities were all decreased in the medium of three day-old cultures grown in the presence of pyrimethanil. The pyrimethanil concentrations resulting in 50% reduction in total enzyme activities (IC₅₀) were approximately 0.25 μM for polygalacturonase, cellulase and proteinase, and approximately 1.0 μM for laccase. No significant growth inhibition was observed at these pyrimethanil concentrations. Pyrimethanil did not inhibit the enzymes directly, nor did it inhibit the synthesis of cytosolic proteins. Therefore, it was proposed that the fungicide inhibits protein secretion at a post-translational stage in the secretory pathway. Large differences were found in the effects of pyrimethanil on the growth of B. cinerea in liquid cultures and on agar plates, depending on the composition of the medium. In liquid media containing cellulose and protein as carbon and nitrogen sources, growth inhibition occurred at 5.0 μM pyrimethanil, whilst no growth inhibition was observed with 50 μM pyrimethanil in malt extract. Similarly, growth occurred on potato/dextrose agar (PDA) at 0.5 μM pyrimethanil, but no growth was seen at this concentration on agars containing cellulose and protein. Thus it appears that pyrimethanil is most active in media where the fungus has to utilise extracellular enzymes to mobilise the nutrients it requires for growth.     

致病杆菌CB43菌株对灰霉病菌的抑制作用

测定了致病杆菌CB43菌株代谢物对灰葡萄病菌菌丝生长和孢子萌发的影响。结果表明:CB43代谢物对灰葡萄孢菌丝生长有较强的抑制作用,500、250、125ml/L代谢物能完全抑制菌丝的生长,并具有杀死作用。62.5ml/L的发酵液处理灰葡萄病菌菌丝72h的抑制率仍达83.35%。20ml/L的发酵液处理灰葡萄孢菌丝能引起菌丝畸形,异常分支、粗短。CB43代谢物能杀死灰葡萄孢分生孢子或抑制其萌发,500~62.5ml/L的发酵液处理16h后,孢子萌发抑制率达88.1%~97.91%。温室试验表明,CB43代谢物对黄瓜灰霉病的控制效果达84.09%,化学农药40%施佳乐的效果为72.94%。