河北省小麦全蚀病菌变种类型鉴定

2007年从河北省的保定、石家庄、邢台小麦主产区采集小麦全蚀病株,共分离得到62个菌株,对其所属的变种类型进行了形态学和分子生物学鉴定。根据形态、培养性状、生理特性以及在小麦、高粱、水稻、玉米、燕麦等禾本科作物上的致病性,初步认定测定的所有菌株均为禾顶囊壳小麦变种(Gaeumannomyces graminisvar.tritici)。进一步采用4个变种的特异性引物进行PCR扩增,在所有菌株中扩增出870bp的条带,该片段为禾顶囊壳小麦变种特异性片段,证实所测菌株均为禾顶囊壳小麦变种。     

河南省小麦全蚀病菌的分离及变种类型鉴定

 从河南省10个地市采集的小麦病根样品中分离得到82株病原分离株, 通过形态特征观察、回接致病性测定、以及分子生物学的方法对所分离菌株进行鉴定。其中47个菌株菌丝分支处都形成典型的“∧”状;子囊壳埋生或半埋生, 子囊棍棒状, 子囊孢子线形, 稍弯曲, 无色, 具有5~10个分隔, 大小为68~94 μm×2~4 μm;对82个分离株rDNA-ITS区进行PCR扩增和序列测定, BLAST序列比对结果表明, 47株菌株与小麦全蚀病菌的ITS序列同源性达97%～99%, 据此确定47株菌株为小麦全蚀病菌。应用小麦全蚀病菌4个变种的特异性引物进行PCR扩增都得到禾顶囊壳小麦变种(870 bp)的特异性片段, 鉴定47株菌株均为禾顶囊壳小麦变种 (Gaeumannomyces graminis var.tritici)。     

小麦全蚀病菌拮抗细菌的筛选及生防效果

为筛选小麦全蚀病菌Gaeumannomyces tritici的拮抗细菌,从健康小麦根际土壤中分离和筛选拮抗细菌,采用对峙培养法和室内盆栽试验测定拮抗细菌对小麦全蚀病菌的抑制活性及生防效果,通过形态学特征观察、生理生化特性测定和保守序列分析确定其分类地位。结果表明,从小麦根际土壤中分离并筛选获得2株具有较好拮抗效果的菌株XJ-3和XJ-4,对小麦全蚀病菌菌丝生长的抑制率分别为64.31%和65.25%。基于菌株XJ-3和XJ-4的形态学特征、生理生化特性和gyrB基因系统进化分析结果,将其均鉴定为贝莱斯芽胞杆菌Bacillus velezensis。菌株XJ-3和XJ-4的发酵液中均含有一类具抗真菌活性、耐热性的非蛋白类次生代谢产物;经菌株XJ-3和XJ-4发酵液处理小麦幼苗后,株高较对照分别显著增加了6.48%和13.12%,干重较对照分别增加了1.30%和14.29%,其中菌株XJ-4的促生效果相对较好;对小麦全蚀病的盆栽防效分别达到51.29%和52.46%。表明筛选得到的贝莱斯芽胞杆菌XJ-3和XJ-4可用于小麦全蚀病的生物防治,具有开发为生防制剂的潜力。     

福建省柑橘褐斑病菌的分离、鉴定和系统进化分析

为明确在福建省南平市的橘柚和三明市的温州蜜橘上发现的疑似柑橘褐斑病的病原菌种类,采用组织分离法获得纯化菌株,通过回接法验证菌株的致病性,利用形态学特征对病原菌进行初步鉴定,并采用最大似然法以多聚半乳糖醛酸酶基因endoPG为靶标对本研究以及国内外已报道的链格孢菌株构建系统发育树,分析其遗传多样性。结果表明,从病组织中共分离获得26株纯培养菌株,经形态学鉴定均为链格孢菌Alternaria spp.。利用分生孢子液接种橘柚离体嫩叶发现,有22株菌株能侵染橘柚叶片并产生与田间相似的褐斑病症状,确认该病害为链格孢引起的柑橘褐斑病。系统发育树分析结果显示,分离所得的26株菌株均聚在已报道的4个柑橘链格孢进化分支Clade1～Clade4中,其中21株菌株聚在国内特有的分支Clade4中,有3株菌株和1株菌株分别聚在国内外兼有的分支Clade3和Clade1中,1株菌株聚在国外特有的分支Clade2中,表明在福建省采集的这些柑橘褐斑病菌均为链格孢菌,且遗传多样性较丰富。     

辣椒轻斑驳病毒凤城分离物的鉴定、全基因测序及系统进化分析

辣椒轻斑驳病毒Pepper mild mottle virus(PMMoV)属于烟草花叶病毒属,是辣椒上重要的病原病毒之一,近年来PMMoV对辽宁部分辣椒产区造成严重危害。本文采用DAS-ELISA检测以及RT-PCR的方法首次从辽宁省凤城市蔬菜产区辣椒病叶中检测出PMMoV,暂命名为PMMoV-FC。设计3对特异性引物对其进行扩增并测序后得到该病毒分离物全基因序列。将PMMoV-FC的全基因序列与已报道的国内外9个PMMoV分离物进行同源性分析,结果表明,其同源性介于94.3%~99.7%之间。基于全基因序列的系统发育进化分析表明,PMMoV-FC与国内分离物、日本分离物以及美洲分离物亲缘关系密切,而与韩国和西班牙分离物亲缘关系稍远。鉴于该病毒在辣椒上造成的严重危害,对于PMMoV-FC在辽宁地区的发生以及防治仍需进行更为详细的研究。     

Molecular phylogenetic analysis reveals at least five genetic races of<Emphasis Type="Italic">Bemisia tabaci</Emphasis> in China

Phylogenetic analysis using ITS1 and CO1 nucleotide sequences has revealed six major races ofBemisia tabaci in the world, including three major indigenous races in the Asia-Pacific region,viz., B. tabaci (Asia),B. tabaci (Bali) andB. tabaci (Australia), but the status of a large collection of genotypes in this region remains unresolved. The ITS1 sequences of representative whitefly samples collected from around China were determined in this study. These sequences and other homologous sequences retrieved from GenBank were then used to conduct a phylogenetic analysis. The results demonstrated that the whiteflies collected in China were split genetically into four groups, where at least five genetic races were revealed,i.e., B biotype (SDLe, XJEp, XJAt, HNNt, BJIb, GDEp, XJGh, GDHrs, XJSm and SHEp), Bali group (ZJGh), M biotype (Hainan1), G biotype (GXCm) and Asian H/K group (FJIb, GDCv), although the Asian H/K group with low bootstrap score remains unresolved. Of all genetic races, the B biotype is the most extensively distributed. In the dendogram, the J biotype, L biotype and Q biotype cluster together and form a sister clade to the B biotype. The data indicate that extensive migration ofB. tabaci has taken place in Asian countries. The populations ZJGh, FJIb, GDCv, GXCm and Hainanl collected in China might have originated there, but the possibility that they were introduced from elsewhere cannot be excluded at this point. Using PyR from Israel as a reference Q biotype, the random amplified polymorphic DNA banding patterns of SDLe, XJEp, XJAt and HNNt were shown to be consistent with that of the Q biotype, which indicated that the four local whitefly populations identified as the B biotype based on ITS1 sequences were closely related to the Q biotype. http//www.phytoparasitica.org posting Sept. 12, 2006.     

一株生防内生真菌的分离筛选、鉴定及抑菌特性

为获取优良植物病害生防菌,以交链孢菌Alternaria tenuis Ness和尖镰孢菌Fusarium oxysporum Schlecht为指示菌,采用平板对峙培养法、抑菌圈法和菌落直径法对健康龙柏鳞叶小枝中分离的4株优势内生真菌进行了筛选评价,并结合形态学观察及ITS序列分析对筛选出的生防内生真菌进行了鉴定。结果表明,对峙培养发现1菌株对交链孢菌和尖镰孢菌均具有较好的抑制作用,抑菌率分别为52.05%和48.65%,标记其为LB-1;LB-1菌株与指示菌对峙培养的菌落交界面处,2种指示菌的菌丝均出现了消解、断裂,而LB-1的菌丝则生长正常。未灭菌LB-1菌株发酵液对交链孢菌和尖镰孢菌产生的抑菌圈直径分别为14.1 mm和13.7 mm,抑菌率分别为47.89%和48.72%;灭菌LB-1菌株发酵液对交链孢菌和尖镰孢菌的抑菌圈直径分别为15.3 mm和15.0 mm,抑菌率分别为5.96%和33.90%,指示菌在生长过程中菌丝溶解现象明显。经鉴定,LB-1菌株为毛壳菌属Chaetomium中的近缘毛壳菌C.subaffine,是一种极具生防潜力的内生真菌,其抑菌特性可能通过溶菌作用实现。     

Effects of powdery mildew and weather on winter wheat yield. 2. Effects of mildew epidemics

Wheat yield losses caused by powdery mildew were computed based on effects of the disease on leaf photosynthesis. Powdery mildew was introduced in a crop model of wheat by quantification of five parameters, taking the vertical and horizontal distribution of mildew in the crop into account. The most important parameters were those of the mildew intensity, the distribution of mildew in the crop, and the effect of mildew on assimilation at light saturation. Measured mildew epidemics in field experiments in three different years, were used to compute yield losses. Computed losses were compared to measured losses. On average, computed yield loss approached measured, but measured yield loss was underestimated, especially in early mildew epidemics due to the computation of partitioning and reallocation of assimilates. Other processes which may cause an underestimation are described. The use of crop models as a method to upgrade disease management systems is discussed.Samenvatting Opbrengstderving van wintertarwe werd berekend aan de hand van het effect dat meeldauw heeft op de blad-fotosynthese. Een rekenmodel voor de gewasgroei van tarwe werd uitgebreid met meeldauw. Met inachtneming van de vertikale en horizontale verdeling van meeldauw in het gewas, werd meeldauw in het model gekwantificeerd door vijf parameters. De belangrijkste parameters waren die van de meeldauwintensiteit, de verdeling van meeldauw in het gewas en het effect van meeldauw op de assimilatie bij een overvloed aan licht. Epidemieën van meeldauw, gemeten in veldproeven in drie verschillende jaren, werden gebruikt om opbrengstdervingen te berekenen. Gemiddeld kwam deze redelijk overeen met de in de veldproeven gemeten opbrengstderving. De gemeten opbrengstderving werd echter onderschat, vooral bij vroege epidemieën van meeldauw door de wijze waarop de (her)verdeling van assimilaten wordt berekend. Andere mechanismen, die een onderschatting van opbrengstderving kunnen veroorzaken worden besproken. Of deze modellen als methode gebruikt kunnen worden om systemen voor de geleide bestrijding van ziekten te verbeteren wordt bediscussieerd.     

Surveys of cereal diseases and pests in the Netherlands. 2. Stem-base diseases of winter wheat

In the period 1974–1986, ca. 100 commercial winter wheat fields were surveyed annually for stembase diseases. In these years, on average 6% of the tillers was infected with eyespot in spring at the first-second, node stage. Eyespot intensity in spring was high in years with high temperature during winter. Eyespot, intensity in the milky-ripe stage in July, averaged 13% of the culms and was high in years with high temperature in April and high precipitation in March, April and May. These data may improve forecasts.In the milky-ripe stage, 2% of the culms were infected with sharp eyespot, but its intensity has gradually increased during the survey years. No significant correlation of sharp eyespot intensity with dry periods in autumn, spring or summer was found.Symptoms of take-all were found on 0.2% of the stem-bases during ripening. Its intensity was low in years with high precipitation in March, April, May and June and high temperature in May and June.Samenvatting Een honderdtal percelen wintertarwe werd in 1974–86 jaarlijks op voetziekten geïnventariseerd. In het voorjaar, tijdens het, eerste en tweede knoopstadium van het gewas, was gemiddeld 6% van de spruiten aangetast door oogvlekkenziekte. De intensiteit van de ziekte was hoog in jaren met een hoge gemiddelde wintertemperatuur. Tijdens het melkrijpe stadium, in juli was gemiddeld 13% van de halmen aangetast door oogvlekkenziekte. De aantasting in juli was hoog in jaren met een hoge temperatuur in april en veel neerslag in de maanden maart, april en mei. Met deze gegevens kunnen adviessystemen worden verbeterd.Scherpe oogvlekkenziekte was op gemiddeld 2% van de halmen in juli aanwezig. De ziekte nam geleidelijk met de jaren toe. De jaarlijkse intensiteit was niet gecorreleerd met droge perioden in de herfst, voorjaar of zomer.Symptomen van halmdoder waren op gemiddeld 0.2% van de halmen aanwezig. De intensiteit van de ziekte was hoog in jaren met weinig neerslag in maart, april, mei en juni en met een lage temperatuur in mei en juni.     

河南省禾谷类作物孢囊线虫的发生分布与种类鉴定

为探究河南省主要禾谷类作物孢囊线虫的发生分布,明确孢囊线虫对不同作物的危害情况,于2017—2020年对河南省18个市50个县（区）的小麦、玉米和水稻作物的孢囊线虫种类和发生分布进行随机取样调查,采用形态学特征、分子生物学鉴定和rDNA-ITS序列进化树分析技术鉴定不同作物孢囊线虫的种类,并根据土壤中孢囊基数和单孢囊卵数明确不同作物孢囊线虫的发病严重度。结果显示,共采集土壤样品308份,其中224份样品检测到孢囊,孢囊检出率为72.7%。小麦孢囊线虫发生分布范围覆盖40个县（区）,其中15个县（区）为禾谷孢囊线虫Heterodera avenae侵染,23个县（区）为菲利普孢囊线虫H. filipjevi侵染,南阳市西峡县西坪镇和开封市尉氏县张市镇为禾谷孢囊线虫和菲利普孢囊线虫混合侵染发生区;玉米孢囊线虫H. zeae在濮阳市清丰县韩村镇、许昌市长葛市董村镇和禹州市范坡镇检测点首次被发现;旱稻孢囊线虫H. elachista在信阳市潢川县魏岗乡、来龙乡和新乡市获嘉县亢村镇检测点首次被发现。孢囊线虫发病严重度数据表明,小麦田平均孢囊含量高达17.3个/100 mL;玉米田平均孢囊量为11.0个/100 mL;水稻田平均孢囊量为4.4个/100 mL。表明河南省孢囊线虫高发地块主要集中在豫北、豫东和豫中平原区。     

陕西省小麦茎基腐病病原菌鉴定及其致病力分析

为明确陕西省小麦茎基腐病病原菌的组成及其致病力,于2019年小麦灌浆期在陕西省宝鸡市、咸阳市、西安市和渭南市共63个采样点采集具有茎基腐病典型症状的病株,对其进行组织分离,结合培养性状和形态学特征,以及利用ITS和EF-1α序列分析鉴定分离菌株所属类群,并对不同病原菌进行致病力分析。结果表明：共分离到224株菌株,基于形态学特征和分子生物学鉴定结果,其中201株菌株鉴定为假禾谷镰刀菌Fusarium pseudograminearum,3株菌株鉴定为禾谷镰刀菌F. graminearum,14株菌株鉴定为三线镰刀菌F. tricinctum,6株菌株鉴定为层出镰刀菌F. proliferatum,其中假禾谷镰刀菌占总分离菌株的89.73%,是陕西省小麦茎基腐病的优势病原菌。不同种的镰刀菌对小麦植株均有一定的致病性,但假禾谷镰刀菌的致病力最强,而且分离自不同地区或者同一地区的假禾谷镰刀菌株间的致病力也存在一定分化。     

Surveys of cereal diseases and pests in the Netherlands. 4. Occurrence of powdery mildew and rusts in winter wheat

Results of annual surveys of winter wheat fields from 1974 to 1986 were compiled to describe epidemics of powdery mildew and rusts in relation to weather and cultivar resistance.An average of 29 and 70% of fields were infected by powdery mildew in May and July, respectively. Mildew prevalence in May was positively correlated with average temperature in October and with average temperature over the months December, January, February and March. In addition, it was correlated negatively with the average grade of mildew resistance of the cultivars sown each year. Prevalence of mildew in July did not show consistent correlations with weather characteristics nor with mildew prevalence in May.Yellow rust was usually not detected in May and on average 18% of the fields was infected in July. The occurrence of yellow rust decreased after 1977, when the farmers adopted cultivars resistant or moderately resistant to yellow rust.Brown rust was usually not detected in May, while in July on average 48% of the fields was infected. Brown rust intensity in July was high in years with a high March temperature and high precipitation during April and May.Black rust was rare in the Netherlands, with 3 and 1% of the fields infected in July 1977 and 1981, respectively.     

山东省小麦根腐病病原菌的分离鉴定

为明确山东省小麦根腐病的病原菌种类,于2012—2014年从山东省10个地市采集小麦病株,通过组织分离法获得了185株分离物,利用形态学鉴定方法,结合基于5.8S r DNA-ITS序列或TEF-1α基因序列分析的分子鉴定方法对分离物进行了鉴定。结果表明:分离物中共得到135株麦根腐平脐蠕孢Bipolaris sorokiniana,占分离病原菌总数的72.97%,属优势种群;50株镰孢属Fusarium菌株,其中14株尖孢镰孢菌Fusarium oxysporum、19株层出镰孢菌Fusarium proliferatum和17株黄色镰孢菌Fusarium culmorum;按照柯赫氏法则进行致病性测定,证实了4种病原菌对鲁麦21号具有致病性,麦根腐平脐蠕孢的致病力较强,病情指数显著高于镰孢菌属真菌。研究表明,山东小麦根腐病主要是由麦根腐平脐蠕孢和镰孢属真菌侵染引起的,麦根腐平脐蠕孢为优势菌群。