Effect of glyphosate on tryptamine production and Sekiguchi lesion formation in rice infected with Magnaporthe grisea

When exposed to light, the Sekiguchi lesion (sl) rice mutant has an enhanced resistance to Magnaporthe grisea infection responsible for Sekiguchi lesion formation and tryptamine accumulation. Glyphosate [N-(phosphonomethyl) glycine] pretreatment suppressed Sekiguchi lesion formation and tryptamine accumulation in the sl mutant after M. grisea infection even under light. This inhibition by glyphosate was blocked by the supply of exogenous tryptophan, but not by exogenous phenylalanine. In glyphosate-pretreated leaves, 5-enol-pyruvyl-shikimate-3-phosphate synthase gene expression and tryptophan biosynthesis were significantly suppressed. During tryptophan starvation, catalase activity was maintained at a high level even under light, leading to the suppression of H₂O₂ generation and DNA fragmentation. These results show a strong relationship between the tryptophan and tryptamine pathways in the induction of light-enhanced resistance to M. grisea infection in the sl mutant.     

Effects of some indole-related compounds on the infection behavior of Magnaporthe grisea

Based on their effect on the infection behavior of Magnaporthe grisea, indole-related compounds were classified into three groups. The first group, including tryptophan, indole-3-butyric acid, indole-3-pyruvic acid, and indole-3-acetamide, did not inhibit infection behavior such as spore germination, appressorium formation, or infection hypha formation in M. grisea. The second, including indole acetic acid, indole-3-acetonitrile, oxindole, and tryptamine inhibited all stages of infection behaviors in a dose-dependent manner. The third, including gramine and indole, did not inhibit spore germination or appressorium formation, whereas it did inhibit infection hypha formation in a dose-dependent manner. These results suggest that endogenous or exogenously applied indole-related compounds in the second and third groups may contribute to protection in blast-susceptible plants such as rice and barley.     

Effect of indole-acetic acid (IAA) on the development of symptoms caused by <Emphasis Type="Italic">Pythium ultimum</Emphasis> on tomato plants

The effect of indole-acetic acid (IAA) on the development of symptoms caused by Pythium ultimum on tomato plants was investigated using different bioassays. Application of IAA (5 μg ml⁻¹) on tomato seedlings inoculated with P. ultimum did not affect their emergence suggesting that IAA did not affect the severity of Pythium damping-off. However, IAA was shown to influence the development of P. ultimum symptoms on tomato plantlets. Low concentrations of IAA (0–0.1 μg ml⁻¹) within the rhizosphere of plantlets increased the severity of the symptoms caused by P. ultimum, while higher concentrations (10 μg ml⁻¹), applied either by drenching to the growing medium or by spraying on the shoot, reduced the symptoms caused by this pathogen. In addition, the study demonstrated that P. ultimum produces IAA in liquid culture amended with L-tryptophan, tryptamine or tryptophol (200 μg ml⁻¹) and in unamended culture.     

Utilization of Parthenium hysterophorus for the remediation of lead‐contaminated soil

A greenhouse experiment was conducted to study the role of plant growth regulators, a chelating agent, and plant growth‐promoting bacteria in lead (Pb) phytoextraction and their subsequent effect on the weed plant, Parthenium hysterophorus. Gibberellic acid (GA₃) and indole‐3‐acetic acid (IAA) were used as the foliar spray. Ethylenediamine tetra acetic acid (EDTA) was applied in split doses. Bacillus and Rhizobium strains were used as a single culture and as co‐cultures. The accumulation of Pb in different parts of the plant was analyzed by using an atomic absorption spectrophotometer. The amount of lead translocation and accumulation in the stems and leaves was significantly higher in the hormonal and EDTA treatments, while the microbial treatments showed no significant difference in the amount of Pb translocation into the stems and leaves, when compared to the control. The EDTA increased the amount of translocation into the shoots, but the dry biomass declined and subsequently reduced the total Pb phytoextraction. The GA₃ treatment showed the maximum total Pb accumulation, along with a higher dry biomass. The microbial co‐inoculated plants showed a significant increase in their dry biomass but the Pb accumulation did not increase like with the GA₃ and IAA treatments. These findings encourage the use of the GA₃ application for Pb phytoextraction by P. hysterophorus. One important feature of this weed plant is its unpalatable nature to herbivores, which could help in reducing the entrance of Pb into the food chain. Gibberellic acid is environmentally friendly compared to EDTA; therefore, more investigation of GA₃ and P. hysterophorus is required.     

干旱胁迫对苗期木薯内源激素含量的影响

以SC124、KU50、C4和SC8四个木薯品种为试验材料,运用酶联免疫吸附测定法,研究了盆栽条件下干旱胁迫对苗期木薯根系与叶片中内源激素脱落酸（ABA）、吲哚乙酸（IAA）、赤霉素（GA）和玉米素核苷（ZR）的含量以及四种激素间平衡关系的影响。结果表明：随着干旱程度的加强,四种激素的含量在不同木薯品种内呈现不同的变化趋势。干旱至第10天时,ABA含量在苗期木薯叶片内增加,在根系内含量也增加（KU50除外）;IAA在叶片中含量下降,根系内含量因品种不同,变化趋势相差明显,其中SC124和KU50中IAA含量显著下降,而在SC8和C4中含量上升;GA含量在四种品种的根系中均下降,叶片GA含量在C4和KU50中略有下降而在SC124和SC8中上升;ZR含量在四种品种的根系中均下降,叶片中ZR含量在C4中上升,SC8中下降,而在KU50和SC124中无明显变化。IAA/ABA ,ZR/ABA在根系中比值下降,SC124降幅最大,SC8降幅最小,KU50、C4降幅居中,说明IAA/ABA ,ZR/ABA与木薯的抗旱能力具有一定相关性。该研究结果解释了苗期木薯根系和叶片内源激素对干旱胁迫的适应性反应,为筛选不同木薯抗旱品种提供了理论依据。     

DNA fragmentation in Sekiguchi lesion mutants of rice infected with Magnaporthe grisea

Light-dependent activation of the tryptamine pathway in Sekiguchi lesion (sl) mutants (Sekiguchi-asahi, Sekiguchi-koshihikari, and Sekiguchi-sasanishiki) inoculated with Magnaporthe grisea, was demonstrated by a significant increase in tryptophan decarboxylase and monoamine oxidase activities and tryptamine accumulation. Terminal deoxynucleotidyl transferase-mediated dUTP-nick end labeling (TUNEL) staining and gel analysis indicated DNA fragmentation in cells of leaf tissues with Sekiguchi lesions of the three sl mutants. Furthermore, increased DNase activity was also light-dependent in the sl mutants after inoculation with M. grisea. DNA fragmentation was inhibited in leaves when Sekiguchi lesion formation was suppressed by cycloheximide and heat shock pretreatments. These data suggest that Sekiguchi lesion formation in the sl mutants is an apoptosis-like response and that its response is induced by light-dependent activation of the tryptamine pathway, which is responsible for light-enhanced resistance.The nucleotide sequence data reported are available in the DDBJ/GenBank/EMBL databases under accession number AB162137 for TDC     

Indole-3-acetic acid biosynthesis in <Emphasis Type="Italic">Aciculosporium take</Emphasis>, a causal agent of witches' broom of bamboo

 Aciculosporium take (Ascomycota; Clavicipitaceae) is a causal agent of witches' broom of bamboo plants. The symptoms of this disease are believed to be induced by plant hormones, particularly auxins. Indole-3-acetic acid (IAA) was identified in cultures of this fungus in an l-tryptophan-supplemented liquid medium. IAA production was confirmed on 30 isolates of A. take from various hosts and locations at levels up to 1 mg/l. The biosynthetic pathway of IAA in A. take culture was examined by analyzing intermediate products and by feeding experiments. The results showed that the indole-3-pyruvic acid pathway (l-tryptophan → indole-3-pyruvic acid → indole acetaldehyde → IAA) was the dominant pathway in A. take. Received: June 3, 2002 / Accepted: July 25, 2002     

Induced resistance against Sclerotinia sclerotiorum in kiwifruit leaves

Salicylic acid (SA), 1-aminocyclopropane-1-carboxylic acid (ACC) and DL -β-amino- n -butyric acid (BABA) were screened for the ability to induce phenylalanine ammonia lyase (PAL) activity in kiwifruit ( Actinidia deliciosa 'Hayward') leaves. SA (2 m M ) was the most effective and induced a 10-fold rise in PAL activity after 2 days compared with a four-fold rise 5 days after ACC (0.5 m M ) treatment. BABA was not an effective elicitor of PAL. SA was further tested, alongside a chlorinated analogue 4-chlorosalicylic acid (4CSA), for the ability to control Sclerotinia sclerotiorum on kiwifruit leaves. Pretreatment with SA and 4CSA caused a reduction in the size of lesions arising from subsequent S. sclerotiorum infection. 4CSA was the more effective and reduced disease levels, relative to controls, by up to 85% on leaf discs and 78% on leaves on the vine. This compares with a 48% reduction by SA on both. Resistance to infection was not affected by washing treated leaf discs prior to inoculation or by delaying inoculation for up to 4 days following 4CSA application. SA and 4CSA were rapidly absorbed and metabolized by kiwifruit leaves and had no apparent phytotoxic effects at the concentrations used for disease control studies. It is proposed that SA and 4CSA operate through the induction of host resistance mechanisms.     

Pathogenicity of indole-3-acetic acid producing fungus <Emphasis Type="Italic">Fusarium delphinoides</Emphasis> strain GPK towards chickpea and pigeon pea

An indole-3-acetic acid (IAA) producing fungal strain was isolated from chickpea grown rhizospheric soil samples. Based on morphological and Internal Transcribed Spacer (ITS) region sequence analysis the new isolate was identified as Fusarium delphinoides. The Fusarium delphinoides strain produces and secretes IAA in-vitro as identified by HPLC and Mass spectrometry. The IAA production is dependent on tryptophan (Trp) as a nitrogen source in the medium. The IAA production is influenced by growth conditions such as pH of the medium, concentration of Trp and the nature of the carbon source. Additional nitrogen sources repress Trp dependent IAA production. Glucose and Trp served as the best carbon and nitrogen sources respectively. Pathogenicity of Fusarium delphinoides towards the plants was tested by electrolyte, nutrient leakage analysis and also by scoring the disease symptoms. Two cultivars of chickpea (ICCV-10 and L-550) and two cultivars of pigeon pea (Maruti and PT-221) were assessed for the pathogenicity by inoculating with spores of Fusarium delphinoides. The inoculation induced symptoms of Fusarium wilt as in the case of Fusarium oxysporum f. sp. ciceris (FOC), a known pathogen causing Fusarium wilt in chickpea. Electrolyte and nutrient leakage from the infected plants were used to assess the resistance, tolerance (moderately resistance) and susceptibility of the plants to the infection. Based on the results, both the pigeon pea cultivars (Maruti and PT-221) were rated as resistant, and ICCV-10 was rated as a tolerant cultivar of chickpea. However, chickpea cultivar L −550 was found to be a susceptible host for infection by Fusarium delphinoides. These results suggest that Fusarium delphinoides, which belongs to the Fusarium dimerum species group, is an IAA producing plant pathogen and causes wilt in chickpea. Further, along with pathogenicity tests, electrolyte and nutrient leakage analysis can be used to assess the pathogenicity of pathogenic fungi.     

Chitosan Stimulates Defense Reactions in Grapevine Leaves and Inhibits Development of Botrytis Cinerea

Chitosan (β-1,4-linked glucosamine oligomer) derived from crab shells conferred a high protection of grapevine leaves against grey mould caused by Botrytis cinerea. Under controlled conditions, it was shown to be an efficient elicitor of some defense reactions in grapevine leaves and to inhibit directly the in vitro development of B. cinerea. Treatment of grapevine leaves by chitosan led to marked induction of lipoxygenase (LOX), phenylalanine ammonia-lyase (PAL) and chitinase activities, three markers of plant defense responses. Dose-response curves show that maximum defense reactions (PAL and chitinase activities) and strong reduction of B. cinerea infection were achieved with 75–150 mg l⁻¹ chitosan. However, greater concentrations of chitosan did not protect grapevine leaves with the same efficiency, but inhibited mycelial growth in vitro. Present results underlined the potency of chitosan in inducing some defense responses in grapevine leaves which in turn might improve resistance to grey mould.     

水杨酸诱导水稻幼苗抗瘟性的生化机制

 以0.01mM水杨酸(SA)喷雾处理或稻瘟菌孢子悬浮液接种稻苗后,叶片中苯丙氨酸解氨酶(PAL)和定位于细胞不同部位的过氧化物酶(POD),特别是与细胞壁结合的POD和细胞间隙POD的活性迅速升高,但没有明显的新POD同功酶带出现。同时叶片中木质素含量迅速增加。在SA处理后的稻叶中提取到能抑制稻瘟菌分生孢子萌发、含有Momilactone A的抑菌物质,并检测到分子量分别为48、52、59、106.5KDa的碱性病程相关蛋白(PRs)。这些生化指标的时序变化与SA诱导抗瘟性的表现相吻合。对SA诱导水稻幼苗抗瘟性的可能生化机理作了归纳。     

AMF+PGPR组合提高黄瓜抗枯萎病的作用机制

对黄瓜(Cucumis sativus)品种香翠16接种尖孢镰刀菌黄瓜专化型(Fusarium oxysporum f.sp.cucumerinum,Foc)、丛枝菌根真菌(AMF)Funneliformis mosseae(Fm)+Foc、丛枝菌根真菌Glomus versiforme(Gv)+Foc、根围促生细菌(PGPR)PS2-6+Foc、PS3-2+Foc、Fm+PS3-2+Foc以及Gv+PS2-6+Foc,以不接种处理作为对照。结果表明,接种Foc条件下,Fm+PS3-2和Gv+PS2-6组合处理的抗病效果显著优于单接种AMF或PGPR处理;镜检观测到靠近AMF菌丝的Foc菌丝纤细,生长不良。AMF菌丝能缠绕Foc菌丝,并阻挡其侵入根系;AMF侵染率与其防效均呈正相关(P0.05);Fm+PS3-2+Foc和Gv+PS2-6+Foc处理的根系活力是单接种Foc的4.6倍;与不接种对照相比,Fm+PS3-2+Foc和Gv+PS2-6+Foc处理的黄瓜叶片中吲哚乙酸(IAA)含量分别提高了205%和166%,赤霉素(GA3)提高了33%和22%,玉米素核甘(ZR)提高了100%和80%,JA平均提高了65%,ABA降低23%和17%;而与单接种Foc处理相比,IAA提高了667%和568%,GA3提高了223%和196%,ZR提高了254%和219%,茉莉酸(JA)含量平均提高了215%,而脱落酸(ABA)含量只有单接种Foc处理的53%;Fm+PS3-2+Foc和Gv+PS2-6+Foc处理显著提高了黄瓜叶内苯丙氨酸解氨酶(PAL)防御酶活性和PAL基因表达量。综上推断AMF+PGPR组合是通过拮抗病原物、促进合成抗病信号物质、上调防御基因表达、提高防御酶活性和降低植株体内有毒物质等机制降低病害和改善植物抗病性。     

Defence responses in flag leaves and spikes of common wheat Triticum aestivum cultivars with contrasting levels of basal resistance to blast caused by Pyricularia oryzae

Wheat blast, caused by Pyricularia oryzae, can cause large yield losses in crops. This study aimed to investigate defence responses in flag leaves and spikes of wheat cultivars BR-18 (moderately resistant) and BRS-Guamirim (susceptible), which differ in their levels of basal resistance. In contrast to cultivar BRS-Guamirim, infected plants of cultivar BR-18 showed more pronounced increases in activities of β-1,3-glucanase and chitinase as well as higher concentrations of lignin-thioglycolic acid derivatives in the flag leaves and total soluble phenolics in the spikes. Polyphenoloxidase activity increased in both flag leaves and spikes in response to fungal infection, regardless of cultivar. Phenylalanine ammonia-lyase (PAL) activity increased in infected flag leaves of both cultivars, especially in BR-18. PAL activity was lower in spikes of infected compared to noninfected plants of both cultivars, although to a lesser extent in BR-18. Compared to BRS-Guamirim, the antioxidative system in both flag leaves and spikes of BR-18 was more efficient in removing reactive oxygen species, reducing cellular damage caused by fungal infection. The lower catalase and peroxidase activities, associated with high superoxide dismutase activity, in flag leaves and spikes of infected BR-18 culminated in a high hydrogen peroxide concentration. The increase in ascorbate peroxidase activity was higher in both flag leaves and spikes of infected plants of BR-18 than in infected BRS-Guamirim. It was concluded that wheat resistance to blast depended on the basal level of resistance of the cultivar, which was mainly associated with the activities of defence enzymes and a more effective antioxidative system.     

Cytological events in tomato leaves inoculated with conidia of <Emphasis Type="Italic">Blumeria graminis</Emphasis> f. sp. <Emphasis Type="Italic">hordei</Emphasis> and <Emphasis Type="Italic">Oidium neolycopersici</Emphasis> KTP-01

Leaves of tomato and barley were inoculated with conidia of Blumeria graminis f. sp. hordei race 1 (R1) or Oidium neolycopersici (KTP-01) to observe cytological responses in search of resistance to powdery mildew. Both conidia formed appressoria at similar rates on tomato or barley leaves, indicating that no resistance was expressed during the prepenetration stage of these fungi. On R1-inoculated tomato leaves, appressoria penetrated the papillae, but subsequent haustorium formation was inhibited by hypersensitive necrosis in the invaded epidermal cells. On the other hand, KTP-01 (pathogenic to tomato leaves) successfully developed functional haustoria in epidermal cells to elongate secondary hyphae, although the hyphal elongation from some conidia was later suppressed by delayed hypersensitive necrosis in some haustorium-harboring epidermal cells. Thus, the present study indicated that the resistance of tomato to powdery mildew fungi was associated with a hypersensitive response in invaded epidermal cells but not the prevention of fungal penetration through host papilla.     

拮抗细菌诱导番茄植株抗灰霉病机理研究

 拮抗细菌多粘类芽孢杆菌(Paenibacillus polymyxa) W3菌株悬浮液及其滤液可以诱导番茄叶片对灰霉病(Botrytis cinerea)的系统抗性。W3及其滤液诱导处理后,植株叶内苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)、多酚氧化酶(PPO)和超氧化物歧化酶(SOD)活性明显增强。诱导后1 d,PAL活性最大,是对照的3.8~3.9倍,6 d后仍为对照的2.5倍;POD和PPO诱导后3 d活性最高,分别比对照增加34.7%~54.1%和78.5%~78.7%,6 d后仍比对照高;SOD活性诱导后2d达高峰,6 d后稍高于对照。活性氧(O₂^-)产生速率诱导后1 d最大,比对照增加85.6%~88.6%,以后急剧下降,6 d后接近对照。此外,W3诱导后1 d或2 d,处理叶和上一叶位叶片水杨酸含量明显上升,分别是对照的2.6倍和1.6倍,这表明该拮抗细菌诱导的系统抗性可能与水杨酸介导有关。     

BION诱导小麦幼苗抗叶锈病研究

研究了BION（有效成分为苯并噻二唑,BTH）对小麦幼苗叶锈病的诱导抗性与苯丙氨酸解氨酶（phenylalanine ammonia lyase,PAL）、多酚氧化酶（polyphenoloxidase,PPO）和过氧化物酶（peroxidase,POD）活性的关系.结果表明：BION诱导麦苗抗叶锈病的最佳浓度为200 mg/L;在诱导处理和接种之间至少需要2天才能诱导抗性表达,间隔4天的诱导抗性效果最好,这种抗性的持久期至少14天.BION处理麦苗第1叶,可使未处理的第2叶也表现出抗性,但处理后至少需要48h这种抗性才能表达.以浓度为200mg/L的BION处理小麦叶片4天后接种,PAL、PPO、POD的活性均高于对照,在接种2天后产生一个峰值,BION水溶液对小麦叶锈菌夏孢子的萌发无明显抑制作用.由此认为,BION可诱导小麦幼苗产生对叶锈病的系统获得抗性.     

Constitutive expression of a phenylalanine ammonia-lyase gene from Stylosanthes humilis in transgenic tobacco leads to enhanced disease resistance but impaired plant growth

Transgenic tobacco plants expressing a phenylalanine ammonia-lyase cDNA (ShPAL), isolated from Stylosanthes humilis, under the control of the 35S promoter of the cauliflower mosaic virus were produced to test the effect of high level PAL expression on disease resistance. The transgenic plants showed up to eight-fold PAL activity and were slowed in growth and flowering relative to non-transgenic controls which have segregated out the transgene. The expression of the ShPAL transgene and elevated PAL levels were correlated and stably inherited. In T₁ and T₂ tobacco plants with increased PAL activity, lesion expansion was significantly reduced by up to 55% on stems inoculated with the Oomycete pathogen Phytophthora parasitica pv. nicotianae. Lesion area was significantly reduced by up to 50% on leaves inoculated with the fungal pathogen Cercospora nicotianae. This study provides further evidence that PAL has a role in plant defence.     

水杨酸对杂交稻抗白叶枯病的作用

杂交稻幼苗感染白叶枯病菌后,其叶片中苯丙氨酸解氨酶(PAL)活性明显提高,丙二醛(MDA)积累和膜透性显著增大,叶绿素含量与不饱和脂肪酸指数(IUFA)下降。经水杨酸(SA)预处理的植株PAL活性仍继续增加,而MDA含量和膜透性的增大趋势得到缓解,膜脂不饱和度增加。作者认为外源SA对杂交稻抗白叶枯病可能有多方面的作用。     

<Emphasis Type="Italic">Pantoea ananatis</Emphasis> strains are differentiated into three groups based on reactions of tobacco and welsh onion and on genetic characteristics

Ninety-six strains of Pantoea ananatis were isolated from 14 plant species including melon, rice, tea and other crops of economic importance. They were classified into three groups (group I, II, III) based on a welsh onion stabbing assay, tobacco infiltration test, and polymerase chain reaction to detect indole acetic acid (IAA) biosynthesis genes (iaaM and iaaH) and a cytokinin biosynthesis gene (etz). Group Ι strains were characterized as causing significant blight symptom on welsh onion and inducing a hypersensitive response (HR)-like reaction on tobacco leaves after 36–48 h and encompassed 20 isolates from foxtail millet, hydrangea, pineapple, river water and rice. These 20 isolates did not possess iaaM, iaaH, or etz genes. Group II, consisting of 34 melon isolates, harbored iaaM, iaaH and etz genes, but did not cause either blight on welsh onion or HR-like reaction on tobacco. Group III strains did not have the iaaM, iaaH, and etz genes, nor did they cause any reaction on welsh onion or tobacco. The 42 strains in group III were isolated from bamboo grass, Chinese silver grass, citrus, dogwood, melon, mugwort, silk tree, sweet corn, tea and welsh onion. Representative strains of the three groups were tested for pathogenicity on melon and rice. Group Ι strains caused palea browning on rice but not internal fruit rot on melon. On the contrary, group II strains did not cause disease on rice but caused internal fruit rot on melon. Group III strains were not pathogenic on rice or melon. These results suggested that the host range of P. ananatis may be predicted by the reactions of welsh onion and tobacco and detection of iaaM, iaaH and etz genes. These tools may serve as rapid tests to identify the pathogenicity groups of P. ananatis.