茄子棒孢叶斑病病原菌鉴定及致病性研究

 本研究对茄子棒孢叶斑病的症状进行系统的描述,采用致病性试验、形态学观察及分子生物学的方法对茄子棒孢叶斑病的病原菌进行鉴定。致病性试验证实病原菌对茄子的致病力显著高于黄瓜。显微观察发现同一病原菌在茄子上形成的孢子以圆柱形为主,孢子体较窄;而在黄瓜上形成的孢子具有圆柱形和倒棍棒形两种形态,且较宽。对病原菌rDNA ITS区进行PCR扩增和序列测定。综合形态特征、致病力和分子序列分析结果,确定茄子棒孢叶斑病病原菌为Corynespora cassiicola。     

The natural occurrence of Thanatephorus cucumeris leaf spot on Hevea brasiliensis in Sri Lanka

Thanatephorus cucumeris. the causal agent of target leaf spot disease of Hevea , is recorded in Sri Lanka for the first time on Hevea brasiliensis .     

Little leaf disease of Piper nigrum in Sri Lanka

In recent years Black pepper ( Piper nigrum ) in Sri Lanka has become affected with'Little leaf'disease. The main symptoms are chlorosis of leaves, shortening of internodes, proliferation of branches, greening and enlargement of floral bracts. Detailed measurements within experimental plots of two improved varieties Panniyur 1 and Kuching, indicated that leaf size, the number of flowers per spike and berry weight were reduced on infected vines. Seeds from infected vines germinated less well than those from healthy vines. In a field trial, disease spread was similar in plots planted with pepper at different spacings and receiving different fertilizer treatments.     

芝麻抗棒孢叶斑病种质资源评价及利用分析

为明确芝麻种质资源对棒孢叶斑病的抗性水平,本试验采用田间人工接菌方法对174份芝麻种质资源抗病性进行评价。田间试验结果表明,供试芝麻材料中没有免疫及高抗品种,通过聚类分析可将供试品种分为5个类群,其中抗病品种32个、中抗品种17个、中感品种69个、感病品种30个、高感品种26个,占参试品种的比例分别为18.39%、9.77%、39.66%、17.24%、14.94%。经亲本系谱分析,抗棒孢叶斑病品种的亲本对棒孢叶斑病也表现良好抗性,说明抗病基因资源的利用对芝麻品种选育至关重要,在育种工作中应加强品种系谱分析,充分利用抗病基因资源。     

番茄棒孢叶斑病病原鉴定及生物学特性研究

本研究采用PSA培养基对上海地区的番茄棒孢叶斑病的病原菌进行分离、培养及致病性测定,通过形态学和分子生物学方法鉴定病原菌,并对该病菌的生物学特性进行研究。结果表明:番茄棒孢叶斑病病原菌菌落呈灰褐色,菌丝质地致密、绒毛状;分生孢子单生或串生,圆柱形或倒棍棒形、顶端钝圆,基部平截,呈半透明至浅褐色,假隔膜4～10个,直或稍弯曲,基脐加厚,深褐色。结合病原菌rDNA ITS序列测定、比对后,确定该病原为多主棒孢Corynespora cassiicola。该病菌的菌丝生长最适温度为25～30℃,最适pH范围为4～8,产孢的最适温度约为25℃,最适pH范围为5～9,麦芽糖和乳糖为碳源的培养基适宜菌丝生长,可溶性淀粉和乳糖溶液中孢子萌发率最高,光暗交替条件适宜菌丝生长,孢子在水滴中极易萌发。这是该病在上海地区的首次报道。     

Comparison of different weed management systems and their effects on yield of coconut plantations in Sri Lanka

The influence of five different weed management systems on nut yield of coconut were evaluated to determine an economical and effective method of controlling weeds in coconut plantations in the low country, dry zone in Sri Lanka. Treatments imposed included slashing and mulching around the palms with slash (T1), slashing and removing the slash (T2), application of glyphosate (N-(phosphonomethyl)-glycine) alone at 1.44 kg ai ha⁻¹ (T3), application of glyphosate alone at 2.88 kg ai ha⁻¹ (T4) and cover cropping with Pueraria phaseoloides (T5). All treatments were applied twice a year, except for the cover cropping treatment, T5. Based on a reduction in weed biomass, treatments T3, T4 and T5 were found to be significantly effective over other treatments. Coconut yield was increased significantly ( P  < 0.05) in glyphosate-applied plots at both tested rates. Control of weeds with the lower concentration of glyphosate (1.44 kg ai ha⁻¹) resulted in a 25% increase in nut yield over the uncontrolled weed plots. At this rate, it was found to be the most effective and economical method of controlling weeds in coconut plantations. Cover cropping with Pueraria phaseoloides was effective in controlling weeds in the long-term, but was not economical compared with the glyphosate application.     

Molecular variability of Apple chlorotic leaf spot virus in Shaanxi,China

Apple chlorotic leaf spot virus (ACLSV) is one of the latent viruses that occur in apple orchards worldwide but usually without visible symptoms. In 2010–2012, a total of 550 apple leaf samples from 12 different major apple-producing areas in Shaanxi, China, were tested by serological assay for ACLSV; the results revealed an infection level of 51.5 %. Because of the known variability in the putative amino acid sequences of the coat protein (CP), and thus the potential for non-detection by serological assay, the molecular variability of isolates of ACLSV collected in Shaanxi was analyzed using PCR and compared with isolates from the rest of the world. Sequences of 504 nt corresponding to 87 % of the CP gene of 12 isolates were acquired by RT-PCR and deposited in GenBank with the accession numbers KF134387–KF134298. Comparisons of the partial CP gene sequences of these 12 isolates as well as isolates previously reported in the world revealed the pairwise identities ranging from 68.9–99.8 % and 73.8–100 % at the nucleotide and amino acid level, respectively. Phylogenetic analysis based on these nucleotide sequences showed that the 72 isolates deposited in GenBank fell into three groups (P205, B6 and Ta Tao 5 Group). Our 12 ACLSV isolates were separated into the P205 and B6 groups, respectively. Multiple alignment analysis of the amino acid sequences of CP revealed that there was a combination of six amino acids at positions 40, 59, 75, 86, 130 and 184 in isolates from each group that could be used to distinguish among the three groups. Two recombination events were identified from all isolates by recombination analysis, and three ACLSV isolates collected in this study participated in these two events. Our results show that molecular variation was present in isolates of ACLSV collected in Shaanxi province and this may reflect introductions of the virus associated with different sources of germplasm.     

来源于新疆的3 个苹果褪绿叶斑病毒分离物的分子变异研究

Apple chlorotic leaf spot virus (ACLSV) isolates from Korla pear (KI-2), New pear no. 7 (XI-1) and Red Fuji apple (API-4) were collected from XinJiang and characterized by analyzing sequences of their near genomic 3忆-terminal. The RT-PCR products were cloned, and analyzed by single-strand conforma-tion polymorphism (SSCP). Eight out of 39 collected positive clones showing different SSCP patterns were sequenced. The results showed that the amplified products had sizes ranging 676 - 703 bp, including partial coat protein (CP) gene (506 bp, accounts for 87% of the complete cp gene) and 3忆-terminal non-coding re-gion (3忆NCR) sequences. The cp gene sequences from isolate KI-2 showed a high intra-isolate divergence,with 84. 8% - 85. 4% identities at the nucleotide (nt) level, and the intra-isolate identities were 99. 8 % and 92.5% - 99. 8 % for isolate XI-1 and API-4, respectively. Phylogenetic analysis on the nt sequences of cpgene showed that the analyzed ACLSV variants from three isolates fell into two different clusters. A variant KI-2-6 from KI-2 was clustered into a group with an apple isolate aclsv-c from China and a plum isolated from France, and all other variants fell into a large cluster. The 3忆NCR sequences of these variants were identical ranging 80. 6% - 100 % .     

解淀粉芽胞杆菌YB114对黄瓜棒孢叶斑病的生防效果及发酵条件优化

解淀粉芽胞杆菌YB114为本研究团队筛选出的一株对黄瓜棒孢叶斑病(Corynespora leaf spot)具有较高活性的生防菌株。为加快其产业化进程,本试验研究了该菌株的生物活性及发酵条件。盆栽试验结果表明,100倍发酵稀释液处理对黄瓜棒孢叶斑病的治疗及预防效果分别为85.72%和42.29%,100倍发酵上清稀释液的治疗及预防效果分别达到74.14%和33.25%,菌株发酵液和上清液对黄瓜棒孢叶斑病的治疗效果好于预防效果。YB114发酵液对黄瓜植株有促生效果,叶面喷施后黄瓜茎长、根长、叶面积及地上部生物量增加,叶片中叶绿素、生长素和玉米核苷素含量显著增加。发酵最优培养基为10 g/L果糖、17 g/L水溶性花生饼粉、1 g/L磷酸二氢钾。最佳发酵条件为：装液量40 mL/250 mL三角瓶,接种量6%(体积分数)、pH 7.0,转速180 r/min,温度32℃,发酵时间24 h。优化发酵条件后YB114对棒孢叶斑病的防效提升了22.4%。     

The detection and characterization of malathion resistance in field populations of Anopheles culicifacies B in Sri Lanka

Malathion resistance was first detected in Sri Lankan Anopheles culicifacies in limited regions of the island in 1982. The frequency of resistance has been increasing slowly since then, but is not yet high enough to be considered an operational problem. Malathion toxicity is synergised in the resistant population by triphenyl phosphate, and metabolism studies suggest the involvement of a carboxylesterase enzyme. The spread of general esterase activity in individuals in an area of the island where resistance is present is wider than that in a totally malathion-susceptible area. However, the frequency of individuals with high esterase activity does not correlate well with resistance in the two field populations studied in detail. This suggests that a qualitative rather than a quantitative change in esterase activity may be involved in this resistance. Extrapolation from similar qualitatively changed carboxylesterases in other anophelines leads us to predict that the resistance in A. culicifacies will be malathion specific and inherited as a single semidominant characteristic.     

云南省玉米灰斑病菌的分子鉴定

本研究利用ITS区、tef1基因、cmdA基因和HIS基因4个基因部分序列对2009-2011年间采自云南省主要灰斑病发生区域和吉林省部分区域的玉米灰斑病菌进行比较分析。结果表明,无论是4个基因序列单独聚类还是拼接序列聚类均把云南省灰斑病发生区域采集的菌株与Cercospora zeina聚为一群,而吉林省采集的灰斑病菌则与Cercospora zeae maydis聚为一群。聚类分析还表明云南省不同玉米产区的灰斑病菌遗传背景单一,在所有聚类树中均聚为一群,且自举支持值均>99%。     

Evidence for oxidative stress involved in physiological leaf spot formation in winter and spring barley

ABSTRACT A leaf spot disease with unknown etiology has become more pronounced in spring and winter barley in Germany in recent years. The symptoms are similar to net blotch and Ramularia leaf spots, but the causal agents of these diseases are not identified. The symptom expression varied much on cultivars. Cultivars most affected by the disease of both spring and winter barley showed a significantly higher level of superoxide (O(2) ) production and lipid peroxidation (malondialdehyde), but a lower level of antioxidant potential expressed as superoxide dismutase (SOD) activity, catalase activity, and integral water-soluble antioxidant capacity (ACW) than insensitive cultivars. A high positive correlation between O(2) production and leaf spot development between ear emergence and milk ripeness was established in the most sensitive winter barley cv. Anoa (r(2) = 0.9622) and spring barley cv. Barke (r(2) = 0.9434). Leaf H(2)O(2) levels increased with the severity of leaf spots. The histochemical localization of O(2) and H(2)O(2) in the tissues adjacent to leaf spots indicated that these two active oxygen species (AOS) are involved in the formation of leaf spots. Reduction of symptom severity by applying strobilurin and azole fungicides was always associated with elevated SOD activity and ACW content and suppressed O(2) production. However, peroxidase activities were significantly higher in sensitive cultivars and in more severely affected tissues and decreased by applying fungicides. Thus, it is assumed that a possible genetic mechanism based on the imbalanced AOS metabolism contributes to formation of physiological leaf spots.     

橡胶狗尾草平脐蠕孢叶斑病病原菌鉴定及生物学特性研究

为明确采自西双版纳景洪地区疑似橡胶麻点病的病原菌,采用组织分离法进行病原菌分离,活体接种法进行致病性测定,通过形态学观察结合rDNA-ITS和GAPDH两个核苷酸片段序列分析,将病原菌确定为狗尾草平脐蠕孢Bipolaris setariae。通过单因子变量试验,对病原菌进行生物学特性研究,结果表明：菌丝生长最适温度为30℃,最适pH为7～8,最适培养基为PDA,菌丝致死温度为53℃,10 min。该病原菌能有效利用多种碳源和氮源,碳源以可溶性淀粉利用率最高,最适氮源为蛋白胨。24 h黑暗条件下培养更有利于菌丝生长。分生孢子萌发的最适温度为28～35℃,最适pH为7～8,28℃黑暗培养2 h后萌发率达到最大,为95.5%,分生孢子致死温度为60℃,10 min。     

Isolation and characterization of rhizobacteria from composts that suppress the severity of bacterial leaf spot of radish

ABSTRACT Composts can induce systemic resistance in plants to disease. Unfortunately, the degree of resistance induced seems highly variable and the basis for this effect is not understood. In this work, only 1 of 79 potting mixes prepared with different batches of mature, stabilized composts produced from several different types of solid wastes suppressed the severity of bacterial leaf spot of radish caused by Xanthomonas campestris pv. armoraciae compared with disease on plants produced in a nonamended sphagnum peat mix. An additional batch of compost-amended mix that had been inoculated with Trichoderma hamatum 382 (T(382)), which is known to induce systemic resistance in plants, also suppressed the disease. A total of 11 out of 538 rhizobacterial strains isolated from roots of radish seedlings grown in these two compostamended mixes that suppressed bacterial leaf spot were able to significantly suppress the severity of this disease when used as inoculum in the compost-amended mixes. The most effective strains were identified as Bacillus sp. based on partial sequencing of 16S rDNA. These strains were significantly less effective in reducing the severity of this disease than T(382). A combined inoculum consisting of T(382) and the most effective rhizobacterial Bacillus strain was less effective than T(382) alone. A drench applied to the potting mix with the systemic acquired resistance-inducing chemical acibenzolar-S-methyl was significantly more effective than T(382) in several, but not all tests. We conclude that systemic suppression of foliar diseases induced by compost amendments is a rare phenomenon. Furthermore, inoculation of compost-amended potting mixes with biocontrol agents such as T(382) that induce systemic resistance in plants can significantly increase the frequency of systemic disease control obtained with natural compost amendments.     

Molecular and serological characterization of iris yellow spot virus, a new and distinct tospovirus species

ABSTRACT A new tospovirus was identified in iris cultivations in the Netherlands. Both serological comparisons and sequence determination of the S RNA demonstrate that this virus represents a new and distinct species, belonging to a separate serogroup, and for which the name iris yellow spot virus (IYSV) is proposed. The disease symptoms on iris are characterized by yellow spots on the leaves. Its experimental host range is very narrow and, in addition to iris, only includes Nicotiana benthamiana and Datura stramonium. The nucleoprotein of IYSV shows only 30 to 44% sequence identity with those of other tospoviruses identified so far; the highest homology being found with the tospovirus species of serogroup IV.     

热带洋兰叶斑病病原菌生物学特性研究和防治药剂筛选

对热带洋兰叶斑病病原菌串珠镰刀茵B10b开展了寄主范围、生物学特性研究,并对其进行了防治药剂毒力测定.结果表明:人工接种后该病原菌能侵染石斛兰、蝴蝶兰、大花蕙兰和文心兰叶片并引起典型叶斑病症状:其菌丝生长的最适温度范围为25～28℃,最适pH7～8:甘露醇、麦芽糖、可溶性淀粉是其最佳生长碳源,牛肉膏和蛋白胨是最适氮源:菌丝和孢子的致死温度为65℃30 min:异菌脲、腈菌唑对菌丝生长的抑制作用很强,其EC50分别为1.12×10-5μg/mL和1.525 4μg/mL,甲基硫菌灵和代森锰锌的效果较弱,其EC50分别为105.879 8μg/mL和66.830 2μg/mL.     

烟草叶斑病病原菌的鉴定及其生物学特性

为明确贵州省新发现的烟草叶斑病病原菌种类和生物学特性,2019年自田间采集具有典型发病症状的烟叶,采用组织分离法对其进行分离纯化,对分离菌株进行致病性测定,观察病原菌形态学特征,基于ITS、LSU、tub2和rpb2基因序列进行分子生物学鉴定,同时对病原菌的生物学特性进行测定。结果显示,烟草叶斑病病斑呈圆形或椭圆形,病斑中间呈浅褐色,边缘呈棕色,周围环绕黄色晕圈。经分离纯化共获得3株病原菌,形成的菌落为深褐色,气生菌丝呈白色;分子孢子器呈球形或扁球形,为深褐色,大小为79.3～91.7μm×148.4～167.3μm;分生孢子呈椭圆形,光滑,无隔膜,具0～3个油球,大小为3.5～5.6μm×1.7～2.9μm。根据形态学特征和分子生物学特征将其鉴定为Didymella segeticola。该病原菌菌丝生长适宜温度为20～25℃;适宜pH为6～10,最适碳源为乳糖,最适氮源为蛋白胨,通气和光暗交替环境有利于其生长。     

来源于桃和苹果的苹果褪绿叶斑病毒的部分分子生物学特性和cp基因的原核表达

 从桃和苹果上分离得到苹果褪绿叶斑病毒ACLSV-HBP和ACLSV-C2个分离物,采用RT-PCR法进行扩增,所获扩增片段经序列测定,其全长分别为1768nt(ACLSV-HBP)和1751nt(ACLSV-C)。这2个分离物扩增片段全长的同源性为83%,mp基因片段核苷酸和推导编码氨基酸序列同源性分别为82.6%和87.1%;cp基因均由582nt组成,其核苷酸和推导编码氨基酸序列同源性分别为87.8%和95.9%。将2个分离物的cp基因与已报道ACLSV分离物进行序列同源性比较,结果显示ACLSV-HBP与SX/2的cp基因核苷酸序列及推导编码氨基酸序列同源性最高,分别为94.0%和96.4%。将ACLSV-HBP分离物的cp基因克隆到原核表达载体pGEX-KG,在大肠杆菌BL21(DE3)中诱导表达,SDS-PAGE分析表明,融合蛋白大小约为46kDa。Western-blot分析表明,该基因在大肠杆菌内得到高效表达,融合蛋白具有抗原性。     

禾草内生真菌与丛枝菌根互作对多年生黑麦草生长及叶斑病的影响

为明确禾草内生真菌和丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)互作对多年生黑麦草Lolium perenne生长发育及叶斑病的影响,设置禾草内生真菌处理(由带有和不带禾草内生真菌种子建立)、AMF(幼套球囊霉Claroideoglomus etunicatum和根内球囊霉Rhizophagus intraradices)单独接种和混合接种处理及不接菌处理(对照),并在植物生长6周后接种或不接种多年生黑麦草叶斑病病原菌根腐离蠕孢Bipolaris sorokiniana,测定各处理多年生黑麦草的发病率、叶绿素含量、净光合速率、水分利用效率、AMF侵染率、P含量和生物量。结果表明:接种病原菌2周后,多年生黑麦草叶斑病的发病率为25.00%～38.75%,禾草内生真菌和幼套球囊霉均在一定程度上降低了多年生黑麦草的发病率,二者共同作用时发病率显著降低了35.48%。禾草内生真菌与AMF互作能在一定程度上提高植物叶绿素含量,促进光合作用,并促进P吸收和植物生长,二者的互作效应因禾草内生真菌与AMF组合而异,但均优于二者单独使用时的效应,其中禾草内生真菌与幼套球囊霉互作对多年生黑麦草生长及叶斑病防治的效果最好。