Hyperparasites of Rhizoctonia solani in Dutch potato fields

Gliocladium roseum was found to be the most common and probably the most effective mycoparasite in potato fields in the northern parts of the Netherlands. It is able to parasitize and kill living hyphae at temperatures of 12°C and higher. Sclerotia ofR. solani are often infected and killed by this fungus under suitable conditions, i.e. at temperatures of 16°C and more. Killing of sclerotia by other antagonistic organisms was also observed. It is also shown by not parasitic fungi and is caused by toxins produced by the antagonist.The development of theG. roseum population was studied during the growth of a potato crop in two soils. In both soils its initial level was very low. In both a slightly acid sandy soil and a neutral sandy loam, suppression ofR. solani can occur;G. roseum accumulated in the former mainly under continuous potato crops,Colletotrichum coccodes was the main antagonist in the latter.Samenvatting In de meeste Nederlandse aardappelakkers komen schimmels voor dieRhizoctonia solani kunnen aantasten en doden. De meest algemene, en waarschijnlijk ook de meest belangrijke, die we tot nu toe vonden, isGliocladium roseum (Tabel 1). Het is bekend, dat deze schimmel stoffen produceert die voorR. solani giftig zijn. Met behulp hiervan kanG. roseum, evenals andere antibiotisch actieve micro-organismen, ook de sclerotiën doden (Tabel 2). Voor doding doorG. roseum is de temperatuur een factor van belang. Hyfen worden nog gedood bij een temperatuur van 12°C, waarbij de sclerotiën niet meer aangetast kunnen worden. Gedurende het winterseizoen worden sclerotiën door deze schimmel naar alle waarschijnlijkheid niet gedood.De ontwikkeling van de populatie vanG. roseum en andere antagonisten vanR. solani werd gevolgd in aardappelvelden op een licht zure zandgrond en op een neutrale zware zavel. Op de zandgrond werden twee proefplekken bemonsterd: één waarop voor het vierde achtereenvolgende jaar aardappelen werden geteeld en één met een vruchtwisselingsschema van graan, bieten en aardappelen.In de zandgrond nam in het groeiseizoen de populatie vanG. roseum toe. Op de proefplek waar voor het vierde jaar achtereen aardappelen stonden werdR. solani vanaf half augustus onderdrukt, evenwel niet volledig. Ook in het vruchtwisselingsstuk breiddeG. roseum zich flink uit, doch een onderdrukking vanR. solani werd niet bereikt.In de zware zavel nam de populatie vanG. roseum niet toe. Hier werdR. solani — uit besmet pootgoed — onderdrukt doorColletotrichum coccodes (zelf een pathogeen van stolonen) en antagonistische bacteriën. De resultaten zijn vermeld in Tabel 3.De besmetting van de geoogste knollen met sclerotiën, zoals die voorkwam op de zandgrond, is in Tabel 4 vermeld. Op de zavel leverde schoon pootgoed een bijna schone oogst (2% van de knollen was zeer licht bezet met sclerotiën). Besmet pootgoed leverde een oogst met 58% schone knollen, 35% met een zeer lichte en 7% met een iets zwaardere sclerotiënbezetting. Hoewel uit 100% besmet pootgoed een veel schonere oogst werd verkregen, was eerder toch een beschadiging van het gewas opgetreden. Pas tegen het eind van het groeiseizoen werdR. solani flink onderdrukt.     

广东省甘薯疮痂病病原菌鉴定及国内主要菜用甘薯种质抗性评价

为明确广东省甘薯疮痂病病原菌种类及当前国内主要菜用甘薯种质对疮痂病的抗性,结合形态学特征和rDNA-ITS序列分析对病原菌进行鉴定,并通过病圃诱发结合人工喷雾接种法对来自9个省区30个菜用甘薯种质进行连续2年的抗性鉴定。结果显示,共获得15株形态学特征及培养性状相似的菌株,其中代表菌株CRI-CJ的形态学特征与甘薯痂圆孢菌Elsinoe batatas基本一致,且在基于rDNA-ITS序列的系统发育树中与甘薯痂圆孢菌聚在一支,表明甘薯疮痂病病原菌为甘薯痂圆孢菌。30个菜用甘薯品种(系)中,抗性、中抗、中感和感病品种(系)分别为7、7、5和11个,占总数的23.3%、23.3%、16.7%和36.7%。其中抗性品种(系)有广菜薯11-52、广菜薯15-6、广菜薯16-1、广菜薯16-19、广菜薯17-23、广菜薯17-9和广菜薯18-6,中抗品种(系)有EC01、广薯菜2号、广菜薯3号、广菜薯6号、广菜薯7号、广菜薯17-25和广菜薯18-3。不同地理来源的品种(系)对甘薯疮痂病的抗性水平存在差异,11个非广东省品种(系)中感病、中感和中抗品种(系)分别为8、2和1个,占比分别为72.7...     

广东省马铃薯块茎软腐病病原菌鉴定

为明确广东省惠州市马铃薯块茎软腐病的病原菌,采用常规病组织分离法获得4株菌株,通过生理生化特征和分子生物学特性对其进行鉴定,并测定了该病原菌对20个马铃薯品种的致病力。结果显示,该病原菌为革兰氏阳性菌,在LB平板上菌落有透明和不透明2种形态;除葡萄糖发酵阳性、对链霉素和青霉素具有抗性等特征不同外,BP-hd-1、BP-hd-2、BP-hd-3和BP-hd-4菌株与短小芽胞杆菌Bacillus pumilus菌株的其它生理生化特征均相同。系统进化分析结果表明,4株菌株的16S rDNA序列分别与短小芽胞杆菌AI-Khrj18(KY123871)、ML270(KC692158)、NCTC10337(LT906438)和ARD21(KX023236)的相似性为100.0%;4株菌株的gyrB基因序列均与短小芽胞杆菌AUEC29菌株(HM585095)的gyrB基因相似性最高,为99.7%～99.8%。生理生化特征及分子生物学鉴定结果表明,引起广东省马铃薯块茎软腐病的病原菌为短小芽胞杆菌。致病力测定结果显示,接种菌株BP-1后20个马铃薯品种的发病率均为100.0%,表明该病原菌对马铃薯有强致病力。     

Mottle necrosis of sweet potato caused by <Emphasis Type="Italic">Pythium scleroteichum</Emphasis> in Japan and varietal difference in susceptibility to the disease

Severe mottle necrosis (Shirogusare-byo in Japanese) was found on mature tubers of sweet potato (Ipomoea batatas) in Ibaraki Prefecture in October 2004. The causal organism was identified as Pythium scleroteichum hitherto unknown in Japan. Sweet potato cultivar Purple Sweet Lord was more susceptible than cultivars Beniazuma, Benimasari, Koukei-14, and Tamayutaka to the pathogen at 25°C, while this difference in the susceptibility was not clear at 15°C.     

取食马铃薯块茎和叶片的马铃薯块茎蛾肠道内可培养细菌的多样性

为明确取食马铃薯块茎和叶片的马铃薯块茎蛾Phthorimaea operculella幼虫肠道可培养细菌种类组成的多样性及其差异,分别对取食马铃薯块茎和叶片的马铃薯块茎蛾4龄幼虫的肠道细菌进行分离培养,根据菌落形态和生理生化特征及16S rDNA序列分析对菌株进行分类鉴定。结果表明,从取食马铃薯块茎的马铃薯块茎蛾肠道内共分离获得细菌有4门8科10属15种,其中褪色沙雷氏菌Serratia marcescen和乙酸钙不动杆菌Acinetobacter calcoaceticus为优势种,其相对多度分别为17.20%和16.06%。从取食马铃薯叶片的马铃薯块茎蛾肠道内共分离获得细菌有4门10科10属16种,其中琥珀葡萄球菌Staphylococcus succinus和乙酸钙不动杆菌为优势种,其相对多度分别为11.86%和15.07%。取食马铃薯块茎和叶片的马铃薯块茎蛾肠道内相同的可培养细菌有4属5种,分别为褪色沙雷氏菌、深红沙雷氏菌Serratia rubidaea、阿氏肠杆菌Enterobacter asburiae、乙酸钙不动杆菌和琥珀葡萄球菌。表明取食马铃薯块茎和叶片的马铃薯块茎蛾...     

河北省甘薯镰孢菌腐烂与溃疡病的病原鉴定

为明确河北省甘薯镰孢菌腐烂与溃疡病的症状特点和病原种类,在不同种植区采集储藏期和育秧期病样,描述其症状特征;对病原菌进行分离纯化,采用柯赫氏法则回接验证,依据病原菌的形态特征和基因序列确定病原菌种类。结果表明,在储藏期甘薯发病可导致薯块表面腐烂的占总病薯的59.09%,端部腐烂的占40.91%;薯块带有褐色轮纹病斑的占61.36%,病斑没有轮纹或者轮纹不明显的占38.64%;发病初期薯块内部病斑浅、黑褐色的占27.27%,发病后期薯块内部形成空腔、布满白色菌丝的占72.73%;病斑带有苦味的占59.09%,病斑没有苦味或苦味不明显的占40.91%;在育秧田发病导致薯秧溃疡,表现为主茎基部呈点片发生黑色或者褐色病斑,部分有开裂。分离的病原菌能够同时侵染薯块和薯秧;病原菌单瓶梗产孢,大型分生孢子稍弯,两端钝圆,多数3~7个分隔,顶细胞钝圆,基细胞足跟较明显。其rDNA-ITS、EF-1α、β-tubulin基因序列与茄镰孢菌Fusarium solani的同源性分别为97%、99%和98%。初步确定甘薯镰孢菌腐烂与溃疡病的病原菌为茄镰孢菌F.solani。     

马铃薯通用应激蛋白StUSP基因家族的鉴定及逆境表达模式分析

在全基因组和转录组水平系统鉴定马铃薯USP通用应激蛋白基因家族,并对该家族成员的基本特征及其在不同逆境胁迫下的表达模式进行分析。同时以马铃薯干旱耐受型品种‘陇薯10号’（L）和干旱敏感型品种‘大西洋’（D）为试验材料,利用高通量测序全面分析StUSP家族成员在干旱胁迫下的表达特性,筛选干旱胁迫关键差异表达基因。结果表明：马铃薯基因组中共有42个基因编码含有USP结构域的蛋白质,在11条染色体上不对称分布,且主要分布在1~6号染色体上。绝大多数StUSPs含有多个内含子。胁迫表达分析显示StUSPs能够响应多种非生物胁迫,且在干旱耐受性不同的马铃薯品种中差异表达。同时,利用qRT-PCR分析了12个成员干旱胁迫下表达模式,除StUSP12、StUSP22下调表达外,其余10个基因均正响应干旱胁迫。     

Identification and management of virulence genes in potato cyst nematodes

Field inoculation of leek with zoospores ofPhytophthora prorri resulted in high infection within a short time. Inoculation with infected leaf tissue resulted in a more gradual increase of disease incidence. Inoculation with oospores was relatively unsuccessful. Zoospores were produced in Petri-dishes by treating fast-growing, young mycelium with a diluted soil extract for at least 2 days, followed by a cold treatment in sterile demineralized water. The successful methods can be used for evaluation of resistance or fungicide performance, and for epidemiological experiments.     

马铃薯丝氨酸羧肽酶类蛋白StSCPL家族的鉴定及干旱胁迫响应分析

在全基因组水平上鉴定了马铃薯SCPL基因家族,并对其成员的基本特征、蛋白质保守结构域、进化关系和干旱胁迫下的表达模式进行了分析。同时,以干旱耐受性不同的马铃薯品种‘陇薯10号’(干旱耐受型)和‘大西洋’(干旱敏感型)为试验材料,利用高通量测序分析了干旱胁迫关键差异表达基因StSCPLs。结果表明,马铃薯中共鉴定到了41个StSCPLs,不对称分布在11条染色体中,多数成员含有多个内含子。其蛋白均含有1个细胞内分泌和转运的信号肽、底物结合位点、N-糖基化位点以及与催化作用相关的进化保守基序。在非生物胁迫下,大多数StSCPLs都具有多胁迫响应性。在干旱耐受性不同的马铃薯品种中,StSCPLs表现出差异表达特性。同时,LncRNA及其靶StSCPL介导的调控网络显示,StSCPL3和StSCPL34分别是lncRNA MSTRG.2301.1和MSTRG.19548.1的靶基因,在干旱耐受性不同的品种中显著差异表达,可能是马铃薯对干旱胁迫响应的潜在靶点。     

宁夏回族自治区马铃薯镰刀菌根腐病病原菌的分离鉴定与致病性测定

为明确宁夏回族自治区马铃薯镰刀菌根腐病的病原菌种类,2015—2018年从宁夏回族自治区固原市、吴忠市、中卫市和石嘴山市4个市12个区县采集马铃薯病样,采用组织分离法对病样进行分离纯化,利用形态学鉴定方法和基于rDNA-ITS与EF-1α序列分析的分子鉴定方法对分离物进行鉴定,并按照柯赫氏法则对其致病性进行测定。结果显示,经组织分离法共获得真菌菌株347株,其中230株镰刀菌属真菌菌株引起马铃薯镰刀菌根腐病,这些镰刀菌分属于5个种,分别为尖孢镰刀菌Fusarium oxysporum、茄病镰刀菌F. solani、锐顶镰刀菌F. acuminatum、木贼镰刀菌F. equiseti、接骨木镰刀菌F. sambucinum,这5种镰刀菌分别为67、29、50、39和45株,分离频率分别为29.13%、12.61%、21.73%、16.95%和19.56%;这5种镰刀菌的致病性有差异,其中接骨木镰刀菌和茄病镰刀菌的发病率分别为91.81%和99.99%,病情指数分别为69.56和82.23,均高于其它3种镰刀菌,是引起宁夏回族自治区马铃薯镰刀菌根腐病的优势病原菌。     

Effect of light duration and growth season on verticillium wilt in potato

The tolerance response to Verticillium in potato is affected by external abiotic and biotic factors. Short-day photoperiod enhanced disease level in the susceptible cvs. ‘Nicola’ and ‘Maris Bard’, as well as in the tolerant cvs. ‘Cara’ and ‘Désirée‘. This was observed under field conditions, where plant reaction in the spring was compared with that in autumn. In some cases the disease was more pronounced in autumn, with its shorter days, than in spring, when the days are longer. The effect was measured by disease symptom severity, fungus colonization in the plants, plant height and final yield. This disease enhancement was more pronounced under controlled growth room conditions, where the only different parameter was illumination. A short photoperiod of 8 h increased the disease level relative to 16 h of light.     

Identification and management of virulence genes in potato cyst nematodes

Efficient and accurate diagnostic assays are essential for the design and evaluation of control measures of the potato cyst nematodesGlobodera rostochiensis andG. pallida by means of resistance. The hybridoma technology and the polymerase chain reaction (PCR) offer in potential various possibilities to design such diagnostic tests for routine purposes. We set out to devise a refined advisory system based on biochemical assays by using the following stepwise approach.In the early 80's a research program was started to develop an immunoassay to differentiate the two sibling species of potato cyst nematodes. Species specific monoclonal antibodies were raised against nematode proteins which are thermostable, abundant and homologous, and which enable reliable species identification using single eggs.     

马铃薯转GhABF2转录因子苗耐盐性研究

以转GhABF2转录因子马铃薯试管苗为材料,进行苗期耐盐性研究,以期了解该作物对逆境的适应性。实验结果表明:在不同浓度NaCl胁迫条件下,转GhABF2基因株系材料(T1、T2)与对照未转基因材料(WT)植株的干鲜重、生理生化指标的变化趋势基本一致,就各性状值的变化来看,转基因植株表现出了明显的抗逆性。随着NaCl胁迫浓度的增加,与对照非转基因植物WT相比,转GhABF2基因材料T1、T2的生物量显著增加;WT、T1、T2的叶绿素含量均随着盐浓度的增加而降低;可溶性糖、丙二醛、脯氨酸,SOD酶活性、POD酶活性均随着盐胁迫浓度的增加而升高;可溶性蛋白含量随着胁迫程度的加重略有下降,但变化趋势不显著。从植株的生长状态、生理生化等性状指标方面来看,转基因植株比对照非转基因植株表现出了更强的抗逆性。     

Density—yield relationships for Colorado potato beetle adults on potatoes

A model composed of two straight lines describes the relation between overwintering spring adults of Colorado potato beetle [Leptinotarsa decemlineata (Say)] and potato yield. Potato vines (Solarium tuberosum (L.), cv. ‘Superior’) had a damage boundary of around 5.8 spring adults per plant. However, under conditions favorable for reproduction, overwintering adults of 0.07 beetles per plant would produce a large population of first-generation larvae that would reduce considerably the potato yield if not controlled. There is no appreciable seasonal overlapping between adults and young larvae in the field. Monitoring activities should therefore be directed at the beginning of the season against overwintering adults and later against first-generation larvae. For summer adults, an action threshold of 10 per plant is proposed.     

马铃薯疮痂病拮抗菌JYC1217的鉴定及其生防促生特性

为获得防治马铃薯疮痂病的优良微生物资源,从健康马铃薯根际土壤中分离获得菌株JYC1217,采用平板对峙培养法和抑菌圈法测定其抑菌活性,结合形态学特征、生理生化特性和分子生物学方法对该菌株进行鉴定,并测定该菌株的生防能力、促生能力及对马铃薯疮痂病的防效。结果显示,菌株JYC1217及其培养液均能显著抑制酸性疮痂链霉菌Streptomyces acidiscabies和肿痂链霉菌S. turgidiscabies的生长;经鉴定菌株JYC1217为多黏类芽胞杆菌Paenibacillus polymyxa;该菌株对马铃薯疮痂病的相对防效为35.12%,对晚疫病菌Phytophthora infestans和早疫病菌Alternaria alternata有抑制作用,抑制率分别为62.96%和64.24%;该菌株能产生β-1,3葡聚糖酶、纤维素酶、蛋白酶和嗜铁素,还可形成复杂生物膜结构,具有固氮、产1-氨基环丙烷-1-羧酸脱氨酶特性,能显著提高马铃薯叶片内苯丙氨酸解氨酶、多酚氧化酶和过氧化物酶活性;该菌株能溶解磷酸钙、大豆卵磷脂和碳酸镁,可显著增加马铃薯株高、叶绿素相对含量、茎粗和单株产量,表明菌株JYC1217是一株生防促生性状优良的生防菌,具有良好的开发应用潜力。     

马铃薯气孔密度表皮模式因子StSTOMAGEN的克隆和功能分析

为了研究马铃薯EPFL9/StSTOMAGEN基因在气孔发育过程中的功能,从马铃薯品种大西洋中克隆得到气孔密度表皮模式因子StSTOMAGEN,将其通过农杆菌介导的遗传转化方法,在拟南芥中异位表达,对该基因的表达特性及功能进行分析。结果显示：StSTOMAGEN主要定位于细胞间隙和细胞核,并且它在顶端未展开叶中表达最多,表达量在1 d中呈现周期性变化,长日照条件下（16 h/8 h,光/暗）,在授时因子为20 h时表达量最高;聚乙二醇（PEG）模拟干旱胁迫处理4 h后其表达量达到最高,而脱落酸（ABA）和NaCl胁迫处理后,其表达水平呈现出持续降低的趋势。功能鉴定表明,过表达StSTOMAGEN基因的拟南芥（ST）相对于野生型（WT）气孔密度增大63%~83%,光合速率增大36%~42%。此外,ST植株离体叶片的水分散失速率增大24%~55%,离体叶片H₂O₂含量也显著升高,约为野生型的50%~100%。干旱胁迫下,ST株系的光合速率、F_v/F_m和瞬时水分利用效率都显著低于野生型,其中光合速率降低55%~66%,F_v/F_m降低22%~50%,瞬时水分利用效率降低11%~12%。综上结果可知,表皮模式因子StSTOMAGEN通过正调控气孔密度而降低植物的抗旱性,该结果为后期通过基因编辑技术降低StSTOMAGEN基因的表达,培育抗旱节水型马铃薯新品种奠定理论基础。     

甘肃定西地区马铃薯线虫病病原的分离鉴定

为明确甘肃省定西市马铃薯线虫病的病原种群分类地位,采用形态学结合分子生物学的方法对该地区马铃薯上的4个线虫群体进行了鉴定,观察和测量其形态特征值,基于r DNA-ITS序列以UPGMA法构建线虫群体的系统发育树,并按照柯赫氏法则进行了致病性测定。结果表明,4个线虫群体在形态学上与马铃薯腐烂茎线虫Ditylenchus destructor一致,但群体DX27与群体DX11、DX16、DX19雌虫的体长、体长/食道长、体长/尾长值存在极显著差异。利用通用引物TW81/AB28扩增r DNA-ITS序列均获得长度为915 bp的片段;序列比对分析表明,群体DX27与其它3个群体相比在ITS1区的第96~255 bp片段内有25个碱基的差异;系统发育树显示,群体DX27与C型群体聚为1支,群体DX11、DX16、DX19与B型群体聚为1支。根据形态学特征及r DNA-ITS序列分析结果确定该病原线虫为马铃薯腐烂茎线虫,其中群体DX27属于C型,群体DX11、DX16、DX19属于B型。     

河北和内蒙古马铃薯干腐病菌种类鉴定

为明确河北和内蒙古马铃薯干腐病菌种类和优势种群,从两省采集300块具干腐病症状的样品,通过分离培养、形态鉴定、TEF-1α序列分析及致病性测定等,对马铃薯干腐病菌种类进行了系统研究.结果表明,两省马铃薯干腐病共存在4种病原菌,即接骨木镰刀菌Fusarium sambucinum、锐顶镰刀菌F.acuminatum、尖孢镰刀菌F.oxysporum和芬芳镰刀菌F.redolens,分别为149、80、3、3株.其中,河北省的优势种群为接骨木镰刀菌和锐顶镰刀菌,其发生频率分别为50％和48.68％;内蒙古的优势种群为接骨木镰刀菌,发生频率为87.95％.     

Molecular identification of some African strains of Ralstonia solanacearum from eucalypt and potato

Ralstonia solanacearum is a known bacterial pathogen of eucalypt and potato plants in Africa. A survey was undertaken to detect this pathogen in eucalypt plantations in South Africa, the Democratic Republic of Congo, and Uganda. Numerous bacterial strains were isolated from trees with symptoms typical of bacterial wilt, but only seven were positively identified as R. solanacearum. A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique, based on the hrp (hypersensitive response and pathogenicity) gene region was used to determine and group the biovars of these R. solanacearum strains. The eucalypt isolates and one potato isolate formed a biovar 3 cluster, whereas the two other potato isolates formed a cluster that corresponded to biovar 2. Amplified fragment length polymorphism (AFLP) analysis confirmed these clusters. Therefore, PCR-RFLP can be used as a reliable diagnostic technique to enable researchers to rapidly identify the pathogen.     

西北地区马铃薯疮痂病病原菌鉴定及其生物学特性

为明确西北地区马铃薯疮痂病病原菌的种类和生物学特性,分别采用常规组织分离法和土壤混悬液分离法从宁夏、陕西和甘肃3个省区采集的29份疮痂病发病薯块和8份发病地块土壤中进行病原菌分离,并利用形态特征、生理生化特性和16S rDNA序列分析对病原菌进行鉴定。结果表明,从发病薯块和发病土壤中共分离到50株链霉菌Streptomyces spp.,通过回接法验证获得6株马铃薯疮痂病致病菌株。6株致病菌株的培养特性和形态特征差别较大;其中菌株G4-1、G9和SYN13不能以果糖和木糖为单一碳源,菌株SYNT3不能以棉子糖为单一碳源;除菌株NLG4-1外,其余5株菌株均能在络氨酸琼脂培养基上产生黑色素。经16S rDNA序列分析,菌株G4-1、G9与疮痂病链霉菌S. scabiei的相似率分别达99.47%和99.34%,菌株NLG4-1、SYNT3与S. enissocaesilis的相似率分别达97.90%和98.18%,菌株GBH2与加利利链霉菌S. galilaeus的相似率达99.93%,菌株SYN13与S. turgidiscabies的相似率达97.56%,表明西北地区马铃薯疮痂病病原菌至少存在4个种。