【Objective】To clarify the pathogen causing Trichosanthes kirilowii fruit rot and screen effective fungicides for controlling the disease.【Method】Samples of rot fruit from T.kirilowii plants were collected from Dabie Mountain Areas,Anhui Province in 2019—2020.A total of 35 representative isolates were obtained by tissue isolation.The pathogenicity was verified according to the Koch’s postulate,and the causal agents were identified based on the morphological characteristics and multilocus sequence analysis.The indoor toxicity of seven fungicides against the pathogen was detected by the mycelium growth rate method.【Result】Colonies with four different morphological characteristics were isolated and purified.The isolates were confirmed to be pathogenic to the fruit of T.kirilowii according to the Koch’s postulate.They were identified as Fusarium fujikuroi,F.proliferatum,Colletotrichum fructicola and C.liaoningense by combining morphological characteristics with phylogenetic analysis,with isolation frequencies of 31.4%,8.6%,20.0% and 40.0% respectively.In addition,the indoor toxicity test results indicated that prochloraz,fludioxonil,difenoconazole phenamacril and chlorothalonil had better inhibitory effect against F.fujikuroi and F.proliferatum,with EC 50 ranging from 0.104 6 to 5.178 1μg/mL.Prochloraz,fludioxonil and difenoconazole had better inhibitory effect against C.fructiicola and C.liaoningense with EC 50 ranging from 0.009 7 to 2.132 5μg/mL.【Conclusion】The causal agents associated with fruit rot disease on T.kirilowii are Fusarium spp.and Colletotrichum spp.Prochloraz,fludioxonil and difenoconazole all show good inhibitory activity against the pathogens.

Maize ear rot, one of the most destructive diseases in summer-sown maize areas of China, is caused by several Fusarium spp. Not only does this pathogen lower maize yields, but also negatively impacts both animal and human health through the production of fumonisin mycotoxins. In this study, we collected 472 unique isolates of F. verticillioides from 480 samples of maize ear rots across eight Chinese summer-sown maize regions and evaluated their genetic variability and aggressiveness. Analysis of polymorphic intersimple sequence repeat (ISSR) loci revealed high levels of genetic variation, particularly in Henan and Shandong provinces. The STRUCTURE result showed that all F. verticillioides isolates clustered into three populations, rather than eight distinct geographical provinces, and there was no significant positive correlation between genetic distance and geographical distance. Most of the variation resulted from genetic differences within rather than between populations. Pathogenicity varied among the evaluated F. verticillioides isolates, and most isolates examined, caused a moderate level of disease; there was no strong correlation between pathogenicity and genetic diversity. This is the first comprehensive report of both the genetic variability and pathogenicity of F. verticillioides across the whole summer-sown maize region in China. These results provide important information that can inform efforts to control maize ear rot and fumonisin contamination caused by F. verticillioides.

BACKGROUND The tea green leafhopper, Empoasca flavescens is the most important pest in Chinese tea plantations. For decades its control has been executed almost exclusively through pesticide applications. A semiochemical-based 'push-pull' strategy was tested on the leafhopper in the study. RESULTS The odors released from Tagetes erecta and Flemingia macrophylla significantly repelled and attracted leafhoppers, respectively. These volatile compounds (46 from T. erecta and 53 F. macrophylla) were identified and quantified via gas chromatography-mass spectometry (GC-MS) analysis. Y-tube olfactometer assays indicated that thymol anisole, thymol and camphor had significant repellent effects on the leafhoppers, resulting in a ternary repellent blend at a 4:3:13 ratio. Cis-3-hexen-1-ol, cis-3-hexenyl acetate, nonanal and alpha-farnesene were significantly attractive to the leafhoppers, making an attractant blend at a 17:4:1:1 ratio. In the field, the push-pull strategy with the repellent dispensers placed within the tea bushes and the attractant-baited sticky traps hung 15 cm above the tea plants showed a significant control efficacy, reaching 69% and 55% at two and 14 days post-treatment, respectively, similar to those in the insecticide control plots. Additionally, the leafhopper density in the push-pull intercropping plot was 63.2 leafhoppers/100 tea shoots/visit, much lower than those in the pull intercropping plot and nonintercropping plot. CONCLUSION Application of the push-pull strategy using both synthetic repellent and attractant, or intercropping T. erecta and F. macrophylla with tea plants, can effectively reduce the leafhopper population. This approach might have great potential as an environmentally safe control strategy against the leafhopper.

丙硫菌唑是目前广泛应用的三唑类广谱杀菌剂.为了明确其对水稻纹枯病菌的生物活性,采用生物测定的方法研究了丙硫菌唑对水稻纹枯病菌的抑菌活性,应用生理生化和分子生物学方法研究了丙硫菌唑对水稻纹枯病菌的可溶性蛋白合成、麦角甾醇合成、DNA合成、菌丝脂质过氧化程度、菌丝细胞膜通透性以及水稻防御酶活性的影响.通过田间药效试验研究了其防治纹枯病效果及对水稻的安全性.结果表明,丙硫菌唑对94株水稻纹枯病菌有较高的抑菌活性,平均EC50值为(1.062 9±0.795 9)mg·L-1.没有出现敏感性下降的抗药性群体,可以采用菌株的平均EC50值作为水稻纹枯病菌对丙硫菌唑的敏感性基线.丙硫菌唑导致病菌麦角甾醇含量降低,菌丝细胞渗透压、菌丝脂质过氧化程度升高,DNA含量和可溶性蛋白含量随降低.丙硫菌唑使POD、CAT、SOD和PAL 4个植物基础防御酶活性升高,具有一定诱导抗病性.田间试验结果显示4.8×105mg·L-1丙硫菌唑悬浮剂,每亩施用剂量为25、30和35mL时,对水稻纹枯病的防治效果分别达90.40％、96.29％和98.17％,与对照试剂噻呋酰胺的防效相当,且对水稻生长安全.

目的 建立QuEChERS-超高效液相色谱-串联质谱法测定葡萄中多菌灵等100种农药及其代谢物的分析方法.方法 样品采用1％乙酸乙腈涡旋振荡提取,离心后取上清液用N-丙基乙二胺(N-propylethylenediamine,PSA)粉末分散固相萃取净化,样品经BEH-C18超高效液相色谱柱分离后,在电喷雾正离子(electrospray ionization,ESI+)模式下,通过多反应监测(multiple reaction monitoring,MRM)模式测定,以基质匹配校准曲线外标法进行定量分析.结果 100种农药及其代谢物在1～100μg/L范围内线性关系良好,相关系数(r2)均大于0.9984;100种农药及代谢物的平均回收率在77.6％～118.4％之间,相对标准偏差为0.3％～28.9％,符合农药多残留分析的要求.结论 本方法简单、灵敏,适用于葡萄中100种农药及其代谢物的准确测定.

为明确安徽省水稻纹枯病的遗传结构及致病分化,采用简单序列重复区间ISSR标记技术和离体叶片接种法对安徽省15个市县的94个水稻纹枯病菌菌株的遗传多样性、遗传结构、致病性及生物学特性进行分析.从18对ISSR引物中筛选出3对重复性好、特异性高的引物对菌株进行PCR扩增,共产生51个ISSR分子标记,其中96.08％的片段具有多态性.安徽省水稻纹枯病菌群体平均有效等位基因数Ne数为1.3138,Shannon信息指数I均值为0.2699,Nei's指数H均值为0.1817,种群基因分化系数平均值为0.4040,表明安徽省水稻纹枯病菌具有丰富的遗传多样性,且不同地理种群间存在较大的遗传分化,分化主要来源于种群内部.UPGMA聚类分析结果表明,同一采集地的菌株大多聚为一类,水稻纹枯病菌种群遗传谱系与其地理来源具有较大的相关性.研究结果为水稻抗纹枯病的种质资源筛选以及抗病品种的合理布局提供了参考.

Recent advances in Colletotrichum taxonomy have led to the need to conduct fresh surveys of Colletotrichum species associated with important crops. Anthracnose caused by Colletotrichum spp. is one of the destructive diseases on Camellia sinensis. In this study, a total of 22 representative Colletotrichum isolates were obtained from diseased leaves of Ca. sinensis cultivated in four tea plantation regions in Anhui Province of China. The isolates were identified based on multilocus (ITS, ACT, CAL, CHS-1, TUB2, GAPDH) phylogenetic analyses, and their morphological characteristics were also analyzed. Twenty-one isolates belonging to C. gloeosporioides complex were identified as C. camelliae, C. fructicola, and C. siamense. One isolate belonging to C. boninense complex was identified as C. karstii. Pathogenicity tests revealed that the isolates of C. camelliae and C. fructicola were highly virulent when inoculated on the leaves of detached twigs of Ca. sinensis cv. Shuchazao. Furthermore, it was found that the interspecies virulence was less distinct and individual isolates showed varied virulence when inoculated on different varieties of Ca. sinensis. To our knowledge, this is the first report of C. fructicola, C. siamense, and C. karstii causing anthracnose on Ca. sinensis in Anhui Province, China.

New fungicides are tools to manage fungal diseases and overcome emerging resistance in fungal pathogens. In this study, a total of 121 Fusarium fujikuroi isolates were collected from various geographical regions of China and their sensitivity to a novel succinate dehydrogenase inhibitor (SDHI) fungicide 'pydiflumetofen' was evaluated. The 50% effective concentration (EC50) value of pydiflumetofen for mycelial growth suppression ranged from 0.0101 to 0.1012 μg/ml and for conidial germination inhibition ranged from 0.0051 to 0.1082 μg/ml. Pydiflumetofen-treated hyphae showed contortion and increased branching, cell membrane permeability, and glycerol content significantly. The result of electron microscope transmission indicated that pydiflumetofen damaged the mycelial cell wall and the cell membrane, and almost broke up the cells, which increased the intracellular plasma leakage. There was no cross-resistance between pydiflumetofen and the widely used fungicides such as carbendazim, prochloraz, and phenamacril. Pydiflumetofen was found safe to seeds and rice seedlings of four rice cultivars, used up to 400 μg/ml. Seed treatment significantly decreased the rate of diseased plants in the greenhouse as well as in field trials in 2017 and 2018. Pydiflumetofen showed superb results against the rice bakanae disease (RBD), when used at 10 or 20 g a.i./100 kg of treated seeds, providing over 90% control efficacy (the maximum control efficacy was up to 97%), which was significantly higher than that of 25% phenamacril (SC) at 10 g or carbendazim at 100 g. Pydiflumetofen is highly effective against F. fujikuroi growth and sporulation as well as RBD in the field.

Meloidogyne graminicola, a devastating plant pathogen of rice (Oryza sativa), is considered to a severe constraint to productivity in rice-growing areas (Zhan et al., 2018). In April 2020, irrigated paddy rice field in Qianshan City, Anhui Province, China, showed symptoms with stunting, thinning, chlorosis, and typical hook-shaped root tips. Females and egg masses of Meloidogyne sp. were found inside the cortex of the root galls, males were found in soil and roots. The morphological characteristics of females, males and second-stage juveniles (J2) were identified as described previously (Tian et al., 2017; Wang et al., 2017). The perineal pattern of the females (n=12) was dorsoventrally oval with low and round dorsal arches, with lateral fields obscure or absent. Most of the striae were smooth, and occasionally broken by short and irregular striae. Morphological measurements (mean±sd, range) of female nematodes (n=20) were body length (543.0±66.0 µm, 448.0-629.0 µm), stylet (11.6±1.9 µm, 7.9-14.2 µm), dorsal pharyngeal gland orifice to stylet base (DGO) (4.0±0.4 µm, 3.4-4.7 µm), vulval slit length (24.1±4.9 µm, 14.8-32.8 µm), vulval slit to anus distance (16.1±3.0 µm, 8.4-20.2 µm). The male nematodes were cylindroid, vermiform, and rounded tail, with the measurements (n=20) body length (1673.0±125 µm, 1346.0-1822.0 µm), stylet (15.5±0.8 µm, 14.0-17.1 µm), DGO (3.7±0.5 µm, 2.9-5.5 µm), and spicule (30.7±2.5 µm, 23.4-34.6 µm). The J2 were vermiform with a gradually tapering hyalines, its measurements (n=20) were body length (452.0±33.0 µm, 391.0-511.0 µm), stylet (13.4±0.8 µm, 12.0-15.2 µm), DGO (3.6±0.6 µm, 2.5-4.7 µm), tail length (72.1±5.2 µm, 59.8-84.8 µm) and hyaline tail terminus (21.7±2.5 µm, 18.0-29.7 µm). DNA extracted from individual females (n=10) were used for molecular identification. The D2/D3 region of 28S RNA was amplified with D2A (5'-ACA AGT ACC GTG AGG GAA AGT TG-3') and D3B (5'-TCG GAA GGA ACC AGC TAC TA-3') (De Ley et al. 1999). The ITS region was amplified with AB28 (5'-ATA TGC TTA AGT TCA GCG GGT-3') and TW81 (5'-GTT TCC GTA GGT GAA CCT GC-3') (Curran et al. 1994). The fragments of D2/D3 region (GenBank accession No. MT576694) and ITS region (GenBank accession No. MT573412) were 766 bp and 579 bp respectively, they all exhibited 99%-100% similarity with those of M. graminicola isolates available in the GenBank. Therefore, both morphological and molecular characterization confirmed the status of this nematode as Meloidogyne graminicola. In green house test, twenty 2-week-old rice seedlings (cv. Longliangyou) were individually maintained in pots with sterilized sand and soil (3:1) and inoculated with 300 J2, other ten non-inoculated rice seedlings as negative control. Rice seedlings were grown in green house at 28℃/25℃ with a 16 h/8 h light/dark photoperiod. After 30 days, all inoculated rice seedling showed symptoms with stunting, chlorosis, and typical hook-shaped root tips, which were similar with that in fields. The nematode reproduction index (final population density/initial population density) were 7.86-10.32. No symptoms were observed on non-inoculated rice seedlings. These results confirmed the pathogenicity of M. graminicola on rice. To our knowledge, this is the first report of M. graminicola in Anhui Province, China. References Curran, J., et al. 1994. Mycol. Res. 98:547. https://doi.org/10.1016/S0953-7562(09)80478-4. De Ley, P., et al. 1999. Nematology. 1:591. https://doi.org/10.1163/156854199508559. Tian, Z., et al. 2017. Plant Disease. http://dx.doi.org/10.1094/PDIS-06-17-0832-PDN. Wang, G., et al. 2017. Plant Disease. http://dx.doi.org/10.1094/PDIS-12-16-1805-PDN. Zhan, L., et al. 2018. BMC Plant Biol. 18:50. https://doi.org/10.1186/s12870-018-1266-9.

为了明确安徽省水稻叶片上发生的一种新病害,从潜山稻田采集病样,利用形态学和分子生物学方法对水稻弯孢叶斑病的病原进行了鉴定,测定了其生物学特性.结果 表明,该病原菌为新月弯孢霉Curvularia lunata,最适于病原菌菌丝生长和产孢的温度为30℃,最适于孢子萌发的温度为25～30℃,致死温度为53℃10 min,最适于菌丝生长、孢子萌发和产孢的pH为6～8,光照对其无影响,最适碳源为蔗糖,最适氮源为铵态氮.

高校教工党支部以政治建设和思想建设铸魂,以组织建设和作风建设强体,以纪律建设以及制度建设立规.不断提高支部政治站位,切实增强支部组织力,突出发挥战斗堡垒作用.教书育人做表率,社会服务当先锋,支部建设树标杆.

为有效防治水稻黑条矮缩病的大面积发生,本研究通过室内大量饲养无毒灰飞虱,饲毒后对浙江省20个主栽水稻品种进行传毒,人工接种水稻黑条矮缩病毒(rice black-streaked dwarf virus,RBSDV)后移栽田间,观察病苗症状,测量株高,统计发病率并分析病毒含量.结果 表明,籼稻品种深两优5814和Y两优302的抗性较强,粳稻品种淮5和秀水14的抗性较弱.此外,病毒侵染后,籼稻品种深两优5814和Y两优302的OsMYC2和OsNPR1的mRNA表达水平显著高于粳稻品种淮5和秀水14.上述结果表明,籼稻品种深两优5814和Y两优302抗病毒能力与其体内茉莉酸(JA)和水杨酸(SA)激素水平相关,这为今后筛选抗RBSDV材料提供了新的策略和思路.

《植物病害流行与预测》是植物保护专业重要的专业基础课程之一.是继《普通植物病理学》和《农业植物病理学》之后开设的提高课程.本文根据植物保护专业人才培养方案,结合《植物病害流行与预测》课程教学的实际,我们课程组从课程目标、课程内容、课程教学及学时分配与课程考核等方面完善并形成了该课程的课程标准;进一步依据该课程质量标准进行了实施运行;讨论与分析了依据课程质量标准实施教学的效果,为相关专业其他课程建设标准的制定提供借鉴.

DNA甲基化作为表观遗传的一种表现形式,在植物的生长发育、抗逆和抗病过程中发挥重要作用.前期研究中从杜仲体内分离筛选到一株拮抗内生芽孢杆菌DZSY21,其可在玉米中定殖,定殖后明显增强玉米抗病性.为明确内生菌DZSY21在玉米中定殖与其DNA甲基化水平的关系,该试验利用甲基敏感扩增多态性分析技术(MSAP)和全基因组DNA甲基化(WGBS)分别对内生菌DZSY21处理的玉米基因组进行DNA甲基化水平的定性和定量研究.定性和定量结果均表明拮抗菌DZSY21引入玉米植株,可引起玉米基因组DNA总甲基化水平降低;全基因组DNA甲基化(WGBS)定量测序结果显示玉米基因组DNA总甲基化水平降低了0.89％,DNA甲基化差异性区域数据库中共有5820个差异性甲基化区域(DMRs),且GO聚类分析和Pathway功能显著性富集分析结果显示DMR相关的基因主要涉及糖代谢、氨基酸合成以及蛋白激酶活性等与抗病相关的主要功能.该结果为深入研究DNA甲基化在内生菌DZSY21定殖增强玉米抗病性中的调控机制提供理论基础.

Pseudomonas fluorescens is a physiologically diverse species of bacteria present in many habitats, which possesses multifunctional traits that provide it with the capability to exhibit biological control activities, promote plant health or cause plant disease. Here, we present the draft genome sequence of the kiwifruit-associated pathogenic isolate AHK-1 of P. fluorescens, which was isolated from the diseased leaves of kiwifruit plants. The genome size of AHK-1 was found to be 7,035,786 bp, with a G + C content of 60.88%. It is predicted to contain a total of 6327 genes, of which 3998 were homologous to genes in the other two sequenced P. fluorescens isolates (SBW25 and GcM5-1A) and 946 were unique to AHK-1 based on comparative genomic analysis. Furthermore, we identified several candidate virulence factors in the genome of AHK-1, including the fliA gene encoding flagellar biosynthetic protein for biosynthesis, and the genes for components of type VI, III, and IV secretion systems. This genomic resource will serve as a reference for better understanding the genetics of pathogenic and non-pathogenic strains, and will help to elucidate the pathogenic mechanisms of P. fluorescens associated with plant disease.

为了筛选防治小麦赤霉病和控制毒素的高效杀菌剂,通过田间小区药效试验及小麦样品DON毒素检测,研究了4种杀菌剂对小麦赤霉病的防治效果和控制毒素的能力.结果表明,36％克菌丹·叶菌唑SC、45％戊唑醇·咪鲜胺EW、48％氰稀·戊唑醇SC和40％戊唑·多菌灵SC在小麦抽穗80％,扬花初期施用,对小麦赤霉病均有较好的防治效果,病指防效在84.25％～93.96％,样品DON毒素检测结果在0.12～0.64μg/ml,其中36％克菌丹·叶菌唑SC 900 ml/hm2的病指防效为93.96％,样品平均DON毒素检测结果为0.12μg/ml,控制毒素效果达93.72％.

明确江西省油菜根肿病病菌类型及油菜品种对病害的抗性,对指导油菜生产及景区观光业的发展具有重要意义.为了探明油菜根肿病的分布及其病菌的致病性分化,于2011年至2016年持续对江西省油菜根肿病的地理分布进行调查,并对病区油菜根肿病株进行生理小种鉴定.结果表明,江西油菜根肿病主要分布在赣东北局部区域,致病菌为4号和9号生理小种,其中4号小种仅发现在赣东北区域分布.在4号和9号小种的病区分别设置田间病圃进行200余个油菜品种的抗性鉴定,结果表明,测试的全部油菜品种对9号小种表现抗病,而94.74％油菜品种对4号小种表现感病;萝卜籽花和59-013等12个品种对4号小种具中抗以上水平.本研究筛选到生理小种特异性的抗病资源或品种,可为油菜抗根肿病育种提供抗源.

根据烟草根黑腐菌与其它近缘真菌在ITS序列上的差异,设计PCR检测引物TbF/TbR,并通过对各菌株基因组DNA扩增验证其特异性;以接种10倍浓度梯度的根黑腐菌孢子的土壤DNA为模板,建立土壤烟草根黑腐菌real-time PCR检测的标准曲线,得出每个反应体系中分生孢子数目的对数(x)和对应的Ct值(y)之间呈线性关系:y=-1.5373x+24.955,R2=0.9909 (P＜0.01).通过对5块烟田土壤烟草根黑腐菌检测和烟田烟草根黑腐病的调查,证实上述方法可以准确检测土壤中烟草根黑腐菌孢子数量.

采集安徽田间典型发病症状的样品,通过组织分离法从无花果病叶和病果上分离获得10个菌株,采用形态学方法和分子生物学方法鉴定并进行致病性测定和柯赫氏法则验证.结果表明,所分离的10个菌株均为胶胞炭疽(Colletotrichum gloeosporioides),其中K7菌株的致病性最强.采用菌丝生长速率法和含毒介质法测定了多菌灵、苯甲·丙环唑、苯甲·嘧菌酯、戊唑醇、 吡唑醚菌酯、 苯醚甲环唑和咪鲜胺等7种杀菌剂对无花果炭疽病菌K7菌株的抑制作用.其中苯丙甲环唑和咪鲜胺对K7菌株的菌丝生长抑制效果最好,EC50值分别为0.0112和0.0130μg/ml.该研究为无花果炭疽病菌的鉴定和高效防治提供了理论依据.

To clarify the effects of Bacillus subtilis BC80-6 on Thielaviopsis basicola, the inhibitory effects of three treatment solutions of B. subtilis BC80-6 on the mycelial growth and spore germination of T. basicola were investigated. BC80-6 fermentation conditions were optimized and field control experiments were carried out. The results showed that 10-1 times living bacteria solution of B. subtilis BC80-6 had the best inhibitory effects on the mycelial growth and spore germination of B. subtilis with the inhibition rates of 93.2% and 95.3% , respectively. Via a single factor test and an orthogonal experiment, the optimum BC80-6 medium was 1.5% sucrose, 0.5% yeast extract and 0.06% magnesium sulfate (mass/volume ratio). The best fermentation condition was fermentation temperature 28 ℃, time 24 h, initial pH 6.5, rotation speed 180 r/min, inoculation amount 5% (v/v) and medium volume 40 mL in 250 mL triangular bottle. The field control effect of living bacteria solution of B. subtilis on tobacco root black rot was 88.19%.