核盘菌Ss160的基因克隆与功能初探

由核盘菌（Sclerotinia sclerotiorum）引起的菌核病是油菜等十字花科作物的重要病害，对油菜的产量品质造成严重影响。本研究基于转录组数据筛选到一个核盘菌基因Ss160 (Sscle04g035160)，发现Ss160在接种油菜的核盘菌中高度表达。生物信息分析表明该基因具有信号肽，且序列在真菌界高度保守。亚细胞定位显示Ss160无特异性的定位，烟草叶片瞬时表达Ss160在核盘菌离体叶接种实验中病斑扩展显著小于表达空载体的和野生型烟草叶片，暗示异源表达Ss160能够提高植物的菌核病抗性。基于酵母系统的体外实验证明Ss160具有转录激活能力。该研究结果为后续Ss160在核盘菌-植物互作中功能机制的研究提供基础，也为油菜菌核病的研究提供借鉴和参考。     

啶酰菌胺与咪鲜胺复配对核盘菌的联合毒力测定

为筛选防治向日葵菌核病的有效药剂,明确不同杀菌剂复配对核盘菌的联合毒力,测定了啶酰菌胺与咪鲜胺复配对核盘菌菌丝生长、菌核的萌发及生理代谢的影响。试验结果表明,不同比例啶·咪复配对核盘菌均有良好的抑制作用,二者复配比例为1:1时,增效系数为1.7657,增效作用最强;啶·咪复配能够有效抑制核盘菌的生长,降低菌核的萌发率和子囊盘形成率,其效果明显优于两单剂;啶·咪复配处理下,核盘菌草酸的分泌量和细胞壁降解酶的活性显著降低,核盘菌的Ss-oah1、Ss-pg1和Ss-pg3等3个致病相关基因的表达量与空白对照相比分别下调了69.90%、55.00%和71.30%。结果表明,啶酰菌胺与咪鲜胺复配,能有效抑制核盘菌的生长,降低核盘菌的致病力,是防治向日葵菌核病的有效复配药剂。     

Relationship between leaf injury and colonization of cabbage by Sclerotinia sclerotiorum

Mechanical and insect feeding injuries of cabbage provided sites for ingress by Sclerotinia sclerotiorum. In field studies, injuries sustained by hitting cabbage heads with a blunt object penetrated several leaf layers and were readily infected by S. sclerotiorum. Incidence of Sclerotinia rot ranged from 15 to 36% infected plants in the hitting treatment. Disease incidence was low (0–5%) when wounds were created by allowing lepidopterous larvae (the imported cabbageworm, Pieris rapae L., the diamondback moth, Plutella xylostella L., and the cabbage looper, Trichoplusia ni Hubner) to feed on cabbage leaves. The incidence of colonization of lepidopterous larvae feeding sites by S. sclerotiorum was greater in greenhouse studies (5–60% infected feeding sites) than in field studies. In greenhouse studies, disease incidence was significantly greater in plants receiving hitting injuries than tearing injuries. There was little evidence of inoculation day or injury day effects. Colonization frequency of tissue injured mechanically was similar whether wounds were inoculated immediately or 4 days later. Plants that were incubated in a dry environment developed fewer infections than plants held in a moist environment. Inoculated plants that were not injured did not develop disease.     

油菜F-box-LRR基因全基因组鉴定与核盘菌诱导应答分析

由核盘菌引起的菌核病是一种重要的真菌性病害,筛选菌核病抗病基因对抗病育种具有重要意义。F-box基因多参与植物抗逆反应,LRR作为抗病基因的重要结构域,在植物抗病防卫中起着重要的作用。本研究通过生物信息学方法在甘蓝型油菜基因组中对F-box-LRR基因进行了全基因组鉴定,基于已发表的中双11组织表达数据以及油菜不同品种中油821（抗病）和Westar（感病）接种核盘菌前后的转录组数据,对可能响应核盘菌诱导的BnF-box-LRR基因进行筛选,并结合荧光定量PCR进行验证。共鉴定到161个BnF-box-LRR基因,从系统进化树上可分为4个亚类（FBXLRR1,FBXLRR2,FBXLRR3和FBXLRR4）,其中第四亚类FBXLRR4在蛋白保守序列分布以及基因结构方面,与其它三个亚类具有较大差异,且与拟南芥参与植物抗逆的同源基因聚为一类,因此推测该分支可能主要参与植物胁迫响应。表达分析表明FBXLRR4家族基因在根和叶中有较高的表达水平,且在核盘菌诱导后具有明显的表达变化,暗示这些基因可能参与油菜菌核病抗性功能。     

Biological control of foliar pathogens by means of Trichoderma harzianum and potential modes of action

Biocontrol of foliar diseases is an alternative means of management of foliar pathogens. One of the most studied commercial biocontrol agents is isolate T39 of Trichoderma harzianum which can be regarded as a model to demonstrate biocontrol under commercial conditions and the mechanisms involved. This biocontrol agent (BCA) controls the foliar pathogens, Botrytis cinerea, Pseuperonospora cubensis, Sclerotinia sclerotiorum and Sphaerotheca fusca (syn. S. fuliginea) in cucumber under commercial greenhouse conditions. Control efficacy was similar for three different rates (covering a fourfold range). Involvement of locally and systemically induced resistance has been demonstrated. Cells of the BCA applied to the roots, and dead cells applied to the leaves of cucumber plants induced control of powdery mildew. The BCA suppressed enzymes of B. cinerea, such as pectinases, cutinase, glucanase and chitinase, through the action of protease secreted on plant surfaces. A combination of several modes of action is responsible for biocontrol. However, biocontrol is not achieved by means of antibiotics or by mycoparasitism, in spite of the fact that BCA has the potential to degrade cell-wall polymers, such as chitin.     

海南百香果病虫害种类调查鉴定及其发生危害

百香果是海南省的新兴产业,也是我国多个省区扶贫攻坚的重要产业。病虫害是影响百香果产业的重要因素,随着海南百香果种植面积越来越大,病虫害对百香果产业的不利影响越来越凸显。由于缺乏百香果病虫害种类调查鉴定的基础性研究,许多种植户对田间发生的病虫害种类识别不足,从而影响了百香果病虫害的防控效果。为了摸清海南百香果种植区的病虫害种类,于2019—2021年对海南省儋州市、乐东县、屯昌县、文昌市、三亚市、东方市、保亭县、白沙县、海口市等百香果种植区的病虫害进行了系统调查与鉴定,并对其发生危害特点进行了研究。鉴定发现海南百香果种植区的病害有11种,分别是病毒病、茎基腐病、炭疽病、疮痂病、疫病、灰霉病、果腐病、褐斑病、烟煤病、藻斑病和缺镁。重要病害为病毒病和茎基腐病,危害程度分别为4级和3级,炭疽病和疮痂病危害也较重,危害程度均为2级,其他病害危害较轻。害虫有28种,分别是黄胸蓟马、茶黄蓟马、红带蓟马、美洲棘蓟马、普通大蓟马、咖啡木蠹蛾、美洲斑潜蝇、橘小实蝇、斜纹夜蛾、双线盗毒蛾、绿丽刺蛾、白带锯蛱蝶、烟粉虱、温室白粉虱、柑橘粉蚧、木槿曼粉蚧、藤壶蜡蚧、茶角盲蝽、樟颈曼盲蝽、稻绿蝽、中稻缘蝽、茶小绿叶蝉、丽球瓢蜡蝉、绿鳞象甲、眼斑芫菁、白条豆芫菁、甘薯梳龟甲和朱砂叶螨。重要害虫为黄胸蓟马和咖啡木蠹蛾,危害程度均为3级,茶黄蓟马和美洲斑潜蝇的危害也较重,危害程度均为2级,其他害虫危害较轻。软体动物3种,分别是灰巴蜗牛、同型巴蜗牛和一种尚未鉴定的蛞蝓。本研究明确了海南百香果种植区病虫害具体种类,新发现百香果病害1种和害虫24种,为海南百香果病虫害的精准防控提供技术支撑。     

Interactions of fungicide-insecticide combinations against Rhizoctonia solani in vitro and in soil

The inhibition of mycelial growth of Rhizoctonia solani in vitro was strongest with pencycuron, followed by tolclofos-methyl, carboxin and thiabendazole. Against cowpea seedling rot in soil infested with R. solani, tolclofos-methyl was most effective, followed by pencycuron, thiabendazole and carboxin. Of nine insecticides tested, only parathion-methyl and quinalphos suppressed mycelial growth of R. solani in vitro, although their activity was much lower than that of the four tested fungicides. The inhibition of mycelial growth by fungicide-insecticide mixtures was antagonistic in only two out of 36 combinations; it was additive in most cases or synergistic, e.g. for most mixtures of pencycuron and insecticides. Quinalphos, applied to the soil, was the only insecticide which reduced cowpea seedling rot in soil infested with R. solani. Soil application of parathion-methyl, phorate, aldicarb or carbofuran and seed treatment with phosphamidon, monocrotophos, endosulfan or dimethoate had little or no effect on seedling rot. Carboxin gave better disease control when applied to the seed already coated with phosphamidon, monocrotophos, endosulfan or dimethoate and when carboxin-treated seeds were sown in soil treated with quinalphos, parathion-methyl, aldicarb or carbofuran. Efficacy of thiabendazole seed treatment was slightly higher in the presence of insecticides, particularly dimethoate. Pencycuron and tolclofos-methyl as seed treatment gave nearly 100% disease control both in the presence and absence of insecticides. The synergistic interactions detected between fungicides and insecticides represent interesting opportunities for the control of R. solani.     

玉米杂交种抗灰斑病鉴定与评价

对目前国内各玉米产区应用的191份玉米杂交种，应用人工注射接种的方法进行抗灰斑病(Cercospora zeae-maydis Tehon & Daiels)鉴定与评价，筛选出对灰斑病表现1级高抗(HR)的杂交种3份、3级抗病(R)的杂交种43份和5级中抗(MR)的杂交种68份。在玉米产区多数杂交种表现为中等抗病或感病，表现高抗的玉米杂交种较少。     

Present state and future prospects of biotechnology in sunflower breeding

The cultivated sunflower is considered to be a species very recalcitrant to various methods of biotechnology, although substantial progress has been made in recent years. For example, very efficient embryo culture techniques are available for obtaining numerous wide crosses which can be very valuable for broadening genetic variation in sunflower. In particular, interspecific hybrids have been demonstrated to be a unique source for creating “new” traits, like cytoplasmic male sterility (CMS) or resistance against devastating diseases, such as Sclerotinia rot. The succesful regeneration of entire plants from cultured somatic tissue or even single cells, i.e. protoplasts, has been demonstrated for specific genotypes. Furthermore, the recovery of androgenetic haploid and doubled haploid plants is basically feasible in sunflower, although the rate of regeneration strongly depends on genotype. Particular progress has been made in the field of “genome characterization”, by using both biochemical and molecular markers. In particular, the chloroplast and mitochondrial genomes have been analyzed and described in detail. Such analyses built the foundation for future “marker-based selection” and for the identification and isolation of specific genes as candidates for “genetic engineering”.     

Occurrence of resistant strains of Botrytis cinerea to anilinopyrimidine fungicides in table grapes in Chile

Effort to control gray mold caused by Botrytis cinerea failed in a table grapevine (Vitis vinifera) vineyard near Santiago, Chile where cyprodinil (Vangard 50 WP), a new fungicide of the anilinopyrimidine group, had been applied alone up to four times per year during two growing seasons. A relatively high frequency (38.5%) of resistant isolates of B. cinerea (EC₅₀ for mycelial growth inhibition varied from 2.9 to 4.84 μg ml⁻¹) may explain the partial loss of field control efficacy obtained. Resistance was correlated with a complete loss of in vivo sensitivity to cyprodinil. Resistant isolates of B. cinerea showed cross resistance to the anilinopyrimidines fungicides mepanipyrim and pyrimethanil. Cyprodinil partially impaired conidia germination and differentially affected conidial germination of resistant and sensitive isolates. Significant differences (p<0.05) in growth rate, sclerotia production and osmotic sensitivity were found among isolates of B. cinerea, but no correlation could be drawn between these biological differences and resistance or sensitivity. This indicates a disruptive selection characteristic of monogenic resistance. Thus, strategies were implemented to avoid the further development and spread of resistance in B. cinerea to the anilinopyrimidine fungicides. To our knowledge this constitutes the first mention of resistance in B. cinerea populations to anilinopyrimidine fungicides in South America.     

钝叶草叶斑病病原鉴定

为鉴定钝叶草叶斑病的病原,通过对61份来自中国海南、广东、广西、云南、福建、江苏,以及美国和南非的疑似钝叶草叶斑病的叶片样品进行分离培养,获得120个菌株,对其进行纯化培养、形态学观察、致病性测定,利用来自海南的钝叶草为靶标寄主,并利用rDNA-ITS、GAPDH、TUB2引物对55号菌株进行分析鉴定。结果表明,经柯赫氏法则验证40株菌株不致病,70株致病性较弱,10株致病性强;选择55号致病性强菌株,进行形态观察,初步确定该病的病原菌为新月弯孢霉（Culvularia lunata）,经3种引物的PCR扩增测序鉴定,和C. lunata相似性达到99%,进一步确定钝叶草叶斑病的致病菌为新月弯孢霉。研究结果将为今后对钝叶草叶斑病的发病规律、防治方法及抗病机制等方面的研究提供理论依据。     

甘蔗黑穗病的防治药剂筛选及产量评价

由甘蔗鞭黑粉菌引起的甘蔗黑穗病是我国甘蔗生产中危害最大的病害,给甘蔗产业造成严重的产量和经济损失。化学防治是作物病害综合防治的重要措施之一,开展甘蔗黑穗病高效、低毒、低残留防治药剂的筛选,进行田间防治试验,对甘蔗黑穗病的有效防治和提高甘蔗产量具有重要意义。为筛选适合大田防治甘蔗黑穗病的推荐化学药剂,通过测定11种杀菌剂对甘蔗鞭黑粉菌的抑菌试验,采用扫描电镜解析有效杀菌剂的抑菌机理,通过新植蔗和宿根蔗的甘蔗黑穗病大田防治试验,评价杀菌剂的防治效果和对产量、株高、茎径、公顷有效茎数、锤度、单茎重等甘蔗主要农艺性状的影响。结果表明,经室内生物活性测定,获得4种杀菌剂对甘蔗鞭黑粉菌单倍体生长、有性配合、冬孢子萌发有抑制作用,分别是42.4%唑醚·氟酰胺悬浮剂、75%肟菌·戊唑醇水分散粒剂、250 g/L吡唑醚菌酯乳油和325 g/L苯甲·嘧菌酯悬浮剂。扫描电镜显示,这4种杀菌剂均对甘蔗鞭黑粉菌冬孢子和单倍体孢子细胞组织造成不同程度的破坏,使甘蔗鞭黑粉菌失活。在大田新植蔗的甘蔗黑穗病防治试验中,42.4%唑醚·氟酰胺悬浮剂、75%肟菌·戊唑醇水分散粒剂、250 g/L吡唑醚菌酯乳油和325 g/L苯甲·嘧菌酯悬浮剂的防治效果分别为91.81%、82.84%、81.76%、70.92%,各药剂防治区产量分别比对照增产20.84%、17.13%、15.92%、13.35%。在宿根蔗的甘蔗黑穗病防治试验中,以上杀菌剂防治效果分别为89.35%、82.21%、81.51%、70.18%,其产量分别比对照增产30.15%、24.05%、20.33%、16.58%。新植蔗和宿根蔗药剂防治区的甘蔗其他农艺性状均优于对照区。这4种杀菌剂对甘蔗黑穗病均有良好的防治效果和增产作用,可作为大田防治甘蔗黑穗病的推荐杀菌剂。     

烟草叶斑病病原菌Epicoccum latusicollum的鉴定、生物学特性及室内药剂筛选

叶斑类病害是危害大田期烟草的主要病害之一，大多由真菌侵染所致，病害易暴发流行，对烟草的品质及产量造成重大影响。本研究采用组织分离法从广西‘K326’烟叶具有典型病斑的叶片上分离、纯化获得菌株，通过致病性测定结合形态学特征以及分子生物学手段对病原菌进行鉴定，同时采用菌丝生长速率法测定不同培养基、温度、pH、碳源、氮源、光照，致死温度处理以及10种常用化学药剂对病原菌菌丝生长的影响。该叶斑病的典型病症为：初期烟叶产生灰白色小圆斑，病害加重圆斑逐渐扩散成不规则状，病斑中心形成穿孔，颜色为灰白色，边缘棕褐色，伴随褪绿的黄色晕圈。通过分离纯化获得2个菌株，分别命名为HZFC36和HZZS76。致病性测定结果表明，2个菌株均可在叶片无伤条件下导致健康‘K326’烟叶部产生病斑。结合形态学特征和多基因位点（LSU、ITS、RPB2和TUB2）系统发育进化分析将引起广西烟草叶斑病的病原菌鉴定为Epicoccum latusicollum。该病原菌在CMA培养基上生长最快，最适生长温度为28℃，最适pH为6，最佳碳源、氮源为蔗糖和牛肉浸粉，全光照条件更利于病原菌的生长，致死温度为47℃，水浴10 min。室内药剂初步筛选结果显示，50%啶酰菌胺和25%吡唑醚菌酯对病原菌的抑制效果最好，平均EC₅₀值分别为4.752×10^-2、4.989×10^-2 mg/L。本研究结果为E. latusicollum引起烟草叶斑病的田间防控奠定了一定的理论基础。     

云南省元江县大麻真菌性病害初步记述

影响元江县大麻产量和质量的主要真菌性病害为菌核病和灰霉病，两种病害导致的损失可达30％以上。其他较常见和导致一定损失的真菌性病害还有秆腐病、霜霉病、黄色叶斑病和褐斑病等。     

油梨根腐病病原菌的鉴定、生物学特性及室内药剂筛选

根腐病是油梨的毁灭性病害之一,该病害为害根部,引起根部变黑腐烂,严重时导致植株死亡。从发病典型的植株根部分离、纯化菌株,并开展致病性测定、形态学特征和多基因位点（ITS、LSU和COXⅡ）系统发育树分析相结合的病原菌鉴定。结果表明,引起海南油梨根腐病的病原菌为樟疫霉（Phytophthora cinnamomi）。还研究了不同培养条件和10种药剂对病原菌菌丝生长的影响。结果表明,病原菌在PDA培养基上生长较好,最适生长温度为28 ℃,最适pH为8,黑暗条件更适合病原菌的生长;烯酰吗啉对病原菌的抑制作用最强,EC₅₀为0.0929 μg/mL。本研究结果为油梨根腐病的田间防控提供了一定的理论依据。     

Steam treatment to prevent carrot decay during storage

Organically grown carrots (Daucus carota L.) have a relatively short storage and shelf life, mainly because chemical measures cannot be implemented to control plant pathogens. Laboratory experiments demonstrated that steam treatments substantially reduced decay of stored organically grown carrots. Prior to being packaged, winter carrots were exposed to 3 s of steam. After 60 days of storage at 0.5°C plus an additional week at shelf conditions (20°C), 2% of the carrots were decayed, as opposed to 23% in the nontreated control. When carrot was inoculated with the fungi Alternaria alternata, [(Fr. : Fr.) Keissler] A. radicina (Meier) and Sclerotinia sclerotiorum [(Lib.) De Bary], percentages of decay, after similar periods of storage and shelf life, were 5% for steam-treated carrot and 65% for the nontreated control. In semi-commercial experiments carrots were treated with steam during the sorting process, and similar results were obtained. Higher decay was found in spring-grown carrots because of the presence of the bacterium Erwinia carotovora.sub. sp. carotovora     

Effects of fungicides on eyespot, caused predominantly by Oculimacula acuformis, and yield of early-drilled winter wheat

The effectiveness against eyespot of nine fungicide treatments, applied at GS 31, was evaluated in early-drilled winter wheat crops. Visual assessments of stem-base diseases were made before fungicide application and at GS 39/55 and 69/70 of cultivars Consort, Savannah and Claire. Competitive PCR was used to quantify Oculimacula acuformis, O. yallundae, Microdochium nivale, Fusarium spp. and Rhizoctonia cerealis. PCR results indicated that O. acuformis was the predominant pathogen causing eyespot in all three crops. Treatments containing cyprodinil were most successful in reducing disease index and DNA of O. acuformis across cultivars. Treatments containing prochloraz were less consistent in eyespot control, significantly reducing the DNA of O. acuformis only in one instance at GS 70 of cultivar Claire. None of the fungicide treatments had any significant effect on brown foot rot or sharp eyespot, or DNA of M. nivale or R. cerealis. The potential for O. acuformis to cause yield loss was indicated by significant negative relationships between yield and eyespot incidence or DNA of O. acuformis at GS 69/70 for each wheat crop. Relationships between DNA of O. acuformis and eyespot incidence or severity at each growth stage were weak but generally improved at the later growth stages of the crops when disease symptoms were easily distinguished. Both molecular methods and visual assessment failed to predict eyespot incidence or pathogen DNA late in season using incidence or DNA of O. acuformis at GS 31. PCR assays, however, accurately identified the stem base pathogens at early growth stages of the crops when the symptoms of their respective diseases were not readily diagnosed.     

Glasshouse and field evaluation of fungicides for the control of Septoria apiicola on celery

Propiconazole was more effective than either triadimenol or hexaconazole when applied after infection, for the control of Septoria apiicola on celery seedlings. In three out of four glasshouse experiments, 25 mg propiconazole I⁻¹ controlled S. apiicola completely when applied within 3 days after inoculation. Propiconazole and triadimenol at 5–15 mg l⁻¹ were also effective when applied 2 days after inoculation. Soil applications of a granular formulation of triadimenol (up to 20 mg per plant) controlled S. apiicola for 5 weeks on celery seedlings grown in pots and, in the field, granules at 100 mg per plant applied at planting were effective for 6 weeks. In two other field experiments the most effective treatments were foliar sprays of 25 mg propiconazole l⁻¹ applied every 7–10 days with or without the addition of anilazine or chlorothalonil.     

Effect of antagonistic Fusarium spp. and of different commercial biofungicide formulations on Fusarium wilt of basil (Ocimum basilicum L.)

The ability of antagonistic Fusarium spp. to control Fusarium wilt of basil caused by F. oxysporum f. sp. basilici was tested under glasshouse conditions from 1994 to 1996. Fusarium oxysporum antagonistic strain 251/2, a protoplast fusion-derived hybrid coded FI-11 and F. moniliforme strains TF4 and TF4 RB were able to decrease (P = 0.05) the incidence of Fusarium wilt significantly. Biocontrol was consistent, especially when the antagonists were applied by mixing a talc preparation of propagules in the substrate (10⁵ CFU/ml of substrate) 1–2 weeks before sowing or at sowing. Seed coating with the biocontrol strains (10⁸ CFU/g of seed) did not generally provide sufficient disease containment, whereas the combination of substrate treatment and seed coating did not improve wilt control in comparison with substrate treatment alone. Two commercial formulations, based on antagonistic F. oxysporum strains and one containing the antagonistic strain K61 of Streptomyces griseoviridis, were generally not sufficiently effective at the tested dosages.     

Impact of varietal resistance in rice and predation on the mortality of Sogatella furcifera (Horváth) (Homoptera: Delphacidae)

Highly resistant, resistant and susceptible rice cultivars were tested in the greenhouse to determine their effect on Sogatella furcifera (Horváth) populations. The highly resistant cultivar IR2035-117-3, with resistance genes Wbph 1 and Wbph 2, caused the highest S. furcifera mortality. When the effects of varietal resistance and predation were combined the increase in mortality due to predators was additive. The spider Lycosa pseudoannulata Boes. et Str., mirid bug Cytorhinus lividipennis Reuter, ladybird beetle Harmonia octomaculata (F.) and rove beetle Paederus fuscipes Curt. were effective predators of S. furcifera and generally caused 30% mortality on the various cultivars. Results indicate that the integration of varietal resistance and predation would provide effective control of S. furcifera populations under field conditions.