白菜叶片挥发成分多样性及对黑斑病菌孢子萌发的影响

本文选取对白菜黑斑病具有不同抗性的白菜品种甜脆绿、瓢儿白、83-1和优早四号,采用GC-MS联用技术对白菜叶面漂洗物的挥发性化学成分进行分析,并用孢子悬滴培养法测定白菜叶面漂洗物对白菜黑斑病菌孢子萌发的影响.结果表明:当叶面漂洗物浓度＞10 μL时,四种白菜叶面漂洗物对病原菌抑制活性的顺序为:瓢儿白＞甜脆绿＞优早四号＞ 83-1,且各品种漂洗物对黑斑病菌孢子萌发的抑制率存在显著差异.甜脆绿漂洗物中含量最高的挥发性化学成分为2,2,4,6,6-五甲基庚烷(17.15％),瓢儿白、83-1和优早四号漂洗物中含量最高的挥发性化学成分为反-3-己烯醇(15.76％、10.87％、7.21％).根据本实验结果发现,2,2,4,6,6-五甲基庚烷、反-3-己烯醇及仅在感病品种83-1漂洗物中检测到的2,4,6,8-四甲基十一烯和2,4,6-ritert-butyl-4-methyl-2,5-cyclo-hexa-dien-1-one可能与黑斑病菌孢子萌发有关.     

Effect of red light and gibberellic acid on lipid metabolism in germinating spores of the fern Anemia phyllitidis

The spores of the fern Anemia phyllitidis contain abundant quantities of lipid as reserve material. Germination of these spores can be induced either by red light or, even in the dark, by gibberellic acid. The effects of both these factors on lipid degradation, lipase and isocitrate lyase activities, and on the fatty acid composition have been studied in the course of the germination process. During germination in darkness with gibberellic acid, the fatty acid composition remained similar to that in the ungerminated spore. In contrast, when spores were germinated in red light, α-linolenic acid was synthesized. Little activity of lipase and isocitrate lyase could be detected in the dry spore. Red light or gibberellic acid affected a dramatic increase of the activities of these enzymes. Lipid breakdown and lipase activity were more active in red light, however. Permanent stimuli were necessary for growth and complete lipid degradation. Induction of germination simultaneously with both factors revealed an additivity of the effects of red light and gibberellic acid.     

甘蓝根肿病菌的生物学特性研究*

对甘蓝根肿病菌生物学特性研究表明,该菌休眠孢子萌发的最适温度24℃,最适pH值6.0～6.7,致死温度45℃,肿根腐烂处理可以显著提高萌发率,光对休眠孢子萌发有明显抑制作用。该菌休眠孢子在感病寄主的根分泌物溶液中萌发率最高,达75%。耐病甘蓝品种及番茄的根分泌物均能刺激休眠孢子萌发。通过电镜观察,根肿病菌休眠孢子为近球形,孢壁有乳状突起,直径2.1～3.1mm(平均直径2.5mm)。游动孢子为近球形或椭圆形,大小为1.6～3.6mm,同侧着生不等长尾鞭式双鞭毛。     

Spore pool glutamic acid as a metabolite in germination

Spore glutamic acid pools were examined in dormant and germinating spores using colorimetric and (14)C analytical procedures. Germination of spores of Bacillus megaterium (parent strain), initiated by d-glucose, was accompanied by a rapid drop in the level of spore pool glutamate, from 12.0 mug/mg of dry spores to 7.7 mug/mg of dry spores after 30 sec of germination. Similar decreases in extractable spore pool glutamate were observed with l-alanine-initiated germination of B. licheniformis spores. On the other hand, glutamate pools of mutant spores of B. megaterium, with a requirement of gamma-aminobutyric acid for spore germination, remained unchanged for 9 min of germination, at which time more than 50% of the spore population had germinated. Evidence for conversion of spore pool glutamate to gamma-aminobutyric acid during germination of spores of B. megaterium (parent strain) was obtained.     

外源茉莉酸诱导植物反应对菜蛾绒茧蜂寄生选择行为的影响

茉莉酸是植物体内重要的伤信号分子,向植物施用外源茉莉酸后, 可诱导植物产生各种防卫反应, 如挥发物组成发生改变等, 进而影响植食性昆虫及其天敌。该文报道用不同浓度外源茉莉酸处理白菜和甘蓝后,诱导植物反应所产生的挥发物对菜蛾绒茧蜂搜索及寄生选择行为的影响。外源茉莉酸处理白菜和甘蓝后,处理植株的挥发物对菜蛾绒茧蜂的引诱力增强;与在对照植株上相比,该蜂对经茉莉酸处理后白菜植株上的小菜蛾幼虫的寄生数显著要高。表明茉莉酸处理白菜及甘蓝后,植物诱导反应导致其挥发物的作用发生变化,进而可提高该蜂的搜索和寄生效率。     

-aminobutyric acid as a required germinant for mutant spores of Bacillus megaterium

Germinated spores of Bacillus megaterium QM B1551 were irradiated with ultraviolet light, and spore-forming survivors were screened for germination requirements. Spore strains which failed to germinate in a variety of defined solutions germinative for spores of the parent strain were obtained. Mutant spores germinated readily in solutions containing yeast extract or one of numerous complex preparations. gamma-Aminobutyric acid, obtained from yeast extract by column chromatography, was shown to be required for germination by the mutant spores. gamma-Aminobutyric acid and l-alanine at final concentrations of 1 mm each, in solutions of KI (40 mm), equaled the potency of yeast extract (1 mg/ml) in the germination of the mutant spores. One of several other amino acids could be substituted, though less effectively, for l-alanine. alpha-Aminobutyric acid, beta-aminobutyric acid, beta-alanine, and 5-aminovaleric acid were ineffective substitutes for gamma-aminobutyric acid in mutant spore germination.     

根际微生态调控白菜根肿病发生的机制研究进展

白菜根肿病是由芸薹根肿菌(Plasmodiophora brassicae Woron)引起的一种常见土传病害,主要危害白菜的根部。根际是土壤-植物-微生物相互作用最活跃的关键微域,根际微生态系统中的微生物失衡是导致土传病害的重要因素,深入探究根际微生态与土传病害互作机制,有利于从根际微生物、抑病物质和功能代谢等方面挖掘防控土传病害安全高效的方法。本文综述了根际微生态与白菜根肿病的发生机制关系,从该病害的危害、发生的根际微生态机制及生防菌防治研究等方面综合分析了根际微生物调控白菜根肿病发生的机制,以期为白菜根肿病防控、促进土壤健康和维持根际微生态系统稳定提供理论依据。     

Role of dipicolinic acid in the germination, stability, and viability of spores of Bacillus subtilis

Spores of Bacillus subtilis spoVF strains that cannot synthesize dipicolinic acid (DPA) but take it up during sporulation were prepared in medium with various DPA concentrations, and the germination and viability of these spores as well as the DPA content in individual spores were measured. Levels of some other small molecules in DPA-less spores were also measured. These studies have allowed the following conclusions. (i) Spores with no DPA or low DPA levels that lack either the cortex-lytic enzyme (CLE) SleB or the receptors that respond to nutrient germinants could be isolated but were unstable and spontaneously initiated early steps in spore germination. (ii) Spores that lacked SleB and nutrient germinant receptors and also had low DPA levels were more stable. (iii) Spontaneous germination of spores with no DPA or low DPA levels was at least in part via activation of SleB. (iv) The other redundant CLE, CwlJ, was activated only by the release of high levels of DPA from spores. (v) Low levels of DPA were sufficient for the viability of spores that lacked most alpha/beta-type small, acid-soluble spore proteins. (vi) DPA levels accumulated in spores prepared in low-DPA-containing media varied greatly between individual spores, in contrast to the presence of more homogeneous DPA levels in individual spores made in media with high DPA concentrations. (vii) At least the great majority of spores of several spoVF strains that contained no DPA also lacked other major spore small molecules and had gone through some of the early reactions in spore germination.     

Antifungal and Biocontrol Evaluation of Four <Emphasis Type="Italic">Lysobacter</Emphasis> Strains Against Clubroot Disease

The effect of crude extract (Ce), seed coating agent (SCA) and whole bacterial broth culture (WBC) of Lysobacter strains was evaluated against the causal agent of clubroot formation in Cruciferous vegetables. The ability of four Lysobacter strains (L. antibioticus 6-B-1, L. antibioticus 6-T-4, L. antibioticus 13-B-1 and L. capsici ZST1-2) inhibited Plasmodiophora brassicae of resting spores and disease. Application of WBC of four Lysobacter strains inhibited clubroot disease, indicating that the disease suppression was due to antifungal compounds produced by the biocontrol bacterium in the culture. Development of clubroot on Chinese cabbage was inhibited when the WBC and SCA were applied before P. brassicae inoculation. Crude extract (Ce) of culture filtrate was effective in arresting the germination of resting spores of P. brassicae on slides. However, Lysobacter strains differed in their biocontrol effects, the strain L. capsci ZST1-2 recorded a high level of disease limiting effect.     

Delayed germination of Bacillus cereus T spores after treatment with trichloroacetic acid and their reactivation by heating

Treatment of Bacillus cereus T spores with trichloroacetic acid delayed their germination. The extent of retardation depended on the concentration of trichloroacetic acid, and the temperature, pH and duration of treatment. The effect was completely reversed by subsequent heating, and this restoration of germination also depended on the temperature and duration of heat treatment. Fourteen compounds were examined for their ability to suppress germination of spores. The halogenated fatty acids tested, such as trifluoro-, tribromo-, and dichloroacetic acid, caused suppression of germination, whereas other compounds, i.e., free fatty acids and amino acids, did not. It is concluded that the charge distribution of fatty acid molecules is important for their effect in suppressing germination of spores.     

Analysis of the action of compounds that inhibit the germination of spores of Bacillus species

AIMS: To determine the mechanism of action of inhibitors of the germination of spores of Bacillus species, and where these inhibitors act in the germination process. METHODS AND RESULTS: Spores of various Bacillus species are significant agents of food spoilage and food-borne disease, and inhibition of spore germination is a potential means of reducing such problems. Germination of the following spores was studied: (i) wild-type B. subtilis spores; (ii) B. subtilis spores with a nutrient receptor variant allowing recognition of a novel germinant; (iii) B. subtilis spores with elevated levels of either the variant nutrient receptor or its wild-type allele; (iv) B. subtilis spores lacking all nutrient receptors and (v) wild-type B. megaterium spores. Spores were germinated with a variety of nutrient germinants, Ca2+-dipicolinic acid (DPA) and dodecylamine for B. subtilis spores, and KBr for B. megaterium spores. Compounds tested as inhibitors of germination included alkyl alcohols, a phenol derivative, a fatty acid, ion channel blockers, enzyme inhibitors and several other compounds. Assays used to assess rates of spore germination monitored: (i) the fall in optical density at 600 nm of spore suspensions; (ii) the release of the dormant spore's large depot of DPA; (iii) hydrolysis of the dormant spore's peptidoglycan cortex and (iv) generation of CFU from spores that lacked all nutrient receptors. The results with B. subtilis spores allowed the assignment of inhibitory compounds into two general groups: (i) those that inhibited the action of, or response to, one nutrient receptor and (ii) those that blocked the action of, or response to, several or all of the nutrient receptors. Some of the compounds in groups 1 and 2 also blocked action of at least one cortex lytic enzyme, however, this does not appear to be the primary site of their action in inhibiting spore germination. The inhibitors had rather different effects on germination of B. subtilis spores with nutrients or non-nutrients, consistent with previous work indicating that germination of B. subtilis spores by non-nutrients does not involve the spore's nutrient receptors. In particular, none of the compounds tested inhibited spore germination with dodecylamine, and only three compounds inhibited Ca2+-DPA germination. In contrast, all compounds had very similar effects on the germination of B. megaterium spores with either glucose or KBr. The effects of the inhibitors tested on spores of both Bacillus species were largely reversible. CONCLUSIONS: This work indicates that inhibitors of B. subtilis spore germination fall into two classes: (i) compounds (most alkyl alcohols, N-ethylmaleimide, nifedipine, phenols, potassium sorbate) that inhibit the action of, or response to, primarily one nutrient receptor and (ii) compounds [amiloride, HgCl2, octanoic acid, octanol, phenylmethylsulphonylfluoride (PMSF), quinine, tetracaine, tosyl-l-arginine methyl ester, trifluoperazine] that inhibit the action of, or response to, several nutrient receptors. Action of these inhibitors, is reversible. The similar effects of inhibitors on B. megaterium spore germination by glucose or KBr indicate that inorganic salts likely trigger germination by activating one or more nutrient receptors. The lack of effect of all inhibitors on dodecylamine germination suggests that this compound stimulates germination by creating channels in the spore's inner membrane allowing DPA release. SIGNIFICANCE AND IMPACT OF THE STUDY: This work provides new insight into the steps in spore germination that are inhibited by various chemicals, and the mechanism of action of these inhibitors. The work also provides new insights into the process of spore germination itself.     

Effects of heat-, CaCl2- and ethanol-treatments on activation of Bacillus spores

The effects of heat, CaCl2, and ethanol on activation of Bacillus spores were determined by monitoring the absorbance decrease during germination in inosine. Bacillus cereus T, B. subtilis A and B. megaterium QM B1551 spores were activated by heat- and CaCl2-treatments. Ethanol activated B. megaterium and B. subtilis spores yet did not activate B. cereus spores. CaCl2- and ethanol-activations were less effective than heat-activation as judged by optimal germination rates and germination extents. The presence of CaCl2 during heat-treatment inhibited heat-activation of all three Bacillus spores without affecting viability or dipicolinic acid content of the spores. The electrophoretic patterns of coat plus outer membrane proteins extracted from Bacillus spores treated with CaCl2 and heat in the presence of CaCl2 were similar to each other and were distinctively different from the patterns of proteins from unactivated spores or the spores treated with heat and/or ethanol.     

Effects of cyanide, salicylhydroxamic acid, and temperature on respiration and germination of spores of the fern Sphaeropteris cooperi

During the first 96 h of culture, germinating spores of the fern Sphaeropteris cooperi (F. v. Muell.) Tryon showed a gradual rise in respiratory activity to a maximum of about 6.5 μl 0₂ h⁻¹ mg⁻¹ dry wt. This was followed by a transitory decline in rate, concluded by a second respiratory rise preceding the emergence of the rhizoid after 192 h of culture. Oxygen uptake during the first 120 h of germination was insensitive to 1 m M potassium cyanide (KCN) but was inhibited by 1 m M salicylhydroxamic acid (SHAM); however, beyond this time cyanide showed increasing inhibitory effectiveness whereas SHAM became less effective. Regardless of time of application, KCN had no effect on germination. Maximum inhibition of germination by SHAM was achieved if applied up to 120 h after culture initiation, after which spores became insensitive to SHAM. Heat treatment (50°C for 90 min) during the cyanide-resistant phase of respiration (0 h–120 h) induced cyanide-sensitive respiration and completely inhibited spore germination. Elevated temperatures had little effect if applied during the cyanide-sensitive phase (beyond 120 h). Temperature inhibited spores regained their ability to germinate if maintained in culture until the cyanide-resistant pathway was restored and then subjected to a second photoinductive light treatment. These results suggest the presence and possible involvement of the cyanide-resistant, alternative respiratory pathway during germination of Sphaeropteris cooperi spores.     

滇重楼种子水浸液对三种植物种子萌发和幼苗生长的影响

利用滇重楼(Paris polyphylla Smith var.yunnanensis(Franch.)Hand.-Mazz.)种子外种皮和胚乳的水浸液对白菜(Brassica pekinensis(Lour.)Rupr.)、绿豆(Vigna radiata(Linn.)Wilczak)、小麦(Triticum aestivum L.)种子进行处理,研究滇重楼种子水浸液对3种植物种子萌发、幼苗生长和保护酶活性的影响,并利用GC-MS方法对滇重楼种子内源抑制物的成分进行分析。结果显示,不同浓度滇重楼外种皮、胚乳水浸液对上述3种受体植物的发芽率、苗高、根长及鲜重均产生影响,其作用强度和水浸液的浓度有关,总体上表现出低促高抑的双重浓度效应。滇重楼种子水浸液对白菜的影响作用最强,对绿豆的影响作用最弱,且胚乳水浸液的影响较外种皮强。不同浓度滇重楼种子外种皮和胚乳水浸液均能影响3种植物幼苗体内保护酶的活性,随着水浸液浓度的升高,叶片中超氧化物歧化酶(SOD)、过氧化物酶(POD)活性总体增加,与对照相比差异显著。白菜、小麦过氧化氢酶(CAT)活性减少,与对照相比差异显著;绿豆过氧化氢酶(CAT)活性增加,但与对照相比无显著差异。利用GC-MS方法从胚乳和外种皮水浸液中分别检出8种和2种物质。研究结果表明滇重楼种子中存在内源抑制物质,可能是导致种子休眠的原因;种子水浸液可能通过影响植物幼苗保护酶的活性进而影响其正常生长;有机酸类物质可能是滇重楼种子内源抑制物之一。     

Germination-induced bioluminescence, a route to determine the inhibitory effect of a combination preservation treatment on bacterial spores

In this work, we have used spores of Bacillus subtilis that specifically induce bioluminescence upon initiation of germination as a rapid, real-time monitor of the effects of preservative treatments on germination. Using this tool, we have demonstrated that the combination of mild acidity (pH 5.5 to 5.0), lactic acid (0. 5%), and a pasteurization step (90 degrees C for 5 min) results in enhanced inhibition of spore germination compared with the effects of the individual treatments alone. Inhibition by the combination treatment occurred as a result of both direct but reversible inhibition, entirely dependent on the physical presence of the preservative factors, and permanent, nonreversible damage to the L-alanine germination apparatus of the spore. However, we were able to restore germination of the preservative-damaged spores unable to germinate on L-alanine by supplementing the medium with the nonnutrient germinant calcium dipicolinic acid. The demonstration that simple combinations of preservative factors inhibit spore germination indicates that food preservation systems providing ambient stability could be designed which do not adhere to the strict limits set by commonly accepted processes and which are based on precise understanding of their inhibitory action.     

Effect of trichloroacetic acid treatment on certain properties of spores of Bacillus cereus T.

Spores of Bacillus cereus T treated with trichloroacetic acid (6.1--61.2 mM) were compared with untreated spores, and as the concentration of the chemical increased, the following alterations in spore properties were found: (1) the extent of germination decreased irrespective of the germination medium used; (2) the spores became sensitive to sodium hydroxide (1 N) and hydrochloric acid (0.27 N), but not to lysozyme (200 micrograms/ml); (3) loss of dipicolinate increased on subsequent heating; and (4) the spores became more sensitive to heat. However, trichloroacetic acid-treated spores were still viable and there was no significant change in spore components. The mechanism of action of trichloroacetic acid is discussed.     

Mechanisms of killing spores of Bacillus subtilis by acid, alkali and ethanol

AIMS: To determine the mechanisms of killing of Bacillus subtilis spores by ethanol or strong acid or alkali. METHODS AND RESULTS: Killing of B. subtilis spores by ethanol or strong acid or alkali was not through DNA damage and the spore coats did not protect spores against these agents. Spores treated with ethanol or acid released their dipicolinic acid (DPA) in parallel with spore killing and the core wet density of ethanol- or acid-killed spores fell to a value close to that for untreated spores lacking DPA. The core regions of spores killed by these two agents were stained by nucleic acid stains that do not penetrate into the core of untreated spores and acid-killed spores appeared to have ruptured. Spores killed by these two agents also did not germinate in nutrient and non-nutrient germinants and were not recovered by lysozyme treatment. Spores killed by alkali did not lose their DPA, did not exhibit a decrease in their core wet density and their cores were not stained by nucleic acid stains. Alkali-killed spores released their DPA upon initiation of spore germination, but did not initiate metabolism and degraded their cortex very poorly. However, spores apparently killed by alkali were recovered by lysozyme treatment. CONCLUSIONS: The data suggest that spore killing by ethanol and strong acid involves the disruption of a spore permeability barrier, while spore killing by strong alkali is due to the inactivation of spore cortex lytic enzymes.SIGNIFICANCE AND IMPACT OF THE STUDY: The results provide further information on the mechanisms of spore killing by various chemicals.     

Germination of surface-disinfected resting spores ofPlasmodiophora brassicae and their root hair infection in turnip hairy roots

Germination of surface-disinfected resting spores ofPlasmodiophora brassicae and its infection of turnip hairy root hairs were studied. Surface-disinfected resting spores showed higher germination than non-disinfected resting spores. Root hair infection was most frequent in the section of root formed 1 d before inoculation. Root hair infection began 4 d after inoculation, increased up to 6 d, and continued to increase more slowly until 10 to 12 d after inoculation. Growth ofP. brassicae in the root hair of hairy roots was observed serially. Most primary plasmodia differentiated to mature zoosporangia 8–10 d after inoculation. The secondary zoospores were initially released 6 d after inoculation.