马铃薯StTCP7基因在干旱和盐胁迫下的表达分析

为了探究马铃薯（Solanum tuberosum L.）TCP7基因在干旱和盐胁迫下的表达模式,以耐旱品种‘Desiree’和干旱敏感品种‘Atlantic’为材料,通过自然干旱和200 mmol·L^-1 NaCl溶液对盆栽苗进行胁迫处理。采用PCR法克隆得到StTCP7基因,CDS区长741 bp,启动子区含有响应干旱诱导的作用元件,蛋白序列与窄刀薯TCP7相似度最高,同源性为98.2%,共编码1个含246个氨基酸的蛋白质;该蛋白含有1个TCP保守结构域,属于碱性蛋白,定位于细胞核上。利用qRT-PCR方法,对干旱和盐胁迫下StTCP7基因在‘Desiree’和‘Atlantic’中的表达量进行分析发现,StTCP7基因表达存在器官差异和品种间差异,在‘Desiree’中的表达量表现为根（1.02）>茎（0.33）>叶（0.29）,在‘Atlantic’中的表达量为叶（4.62）>茎（3.72）>根（1.00）。干旱胁迫下,StTCP7基因在‘Desiree’根和叶中的表达量随干旱胁迫程度的增加呈先上升后下降趋势,W3（土壤含水量为35%~45%）时达到峰值,其表达量分别为对照组的1.27倍和1.87倍;该基因在‘Atlantic’根部的表达量呈先上升后下降的趋势,W3（35%~45%）时达到峰值,其表达量为对照组的1.39倍,叶中表达量呈持续上升趋势,W4（15%~25%）时达到峰值,其表达量为对照组的2.52倍。盐处理下,随胁迫时间的增加,StTCP7基因在‘Desiree’根部的表达量呈先上升后下降的趋势,处理6 h时达到最高,为对照组的2.16倍;叶中表达量呈先上升后下降的趋势,处理3 h时达到最高,为对照组的4.54倍;该基因在‘Atlantic’根部的表达量呈现先上升后下降的趋势,胁迫3 h时达到最高,为对照组的2.12倍,叶中表达量呈上升趋势,在48 h时达到峰值,其表达量为对照组的4.04倍。综上所述,马铃薯StTCP7基因响应干旱与盐胁迫,且在根和叶不同器官中的表达存在差异,可为StTCP7基因在马铃薯非生物逆境响应中的功能研究提供基础。     

白菜型油菜BraSAUR基因家族鉴定及其表达分析

SAUR(Small auxin-up RNA)是生长素早期响应逆境胁迫的基因,本文研究白菜型油菜全基因组中SAUR家族的信息,分析了该家族成员的基本特征。以强抗寒性白菜型冬油菜品种陇油7号（L7）和弱抗寒性冬油菜品种陇Lenox（X）为试验材料,采用低温和干旱处理,应用荧光定量技术分析不同基因在不同品种中的表达特性,筛选差异表达基因,为研究生长素早期应答基因调控白菜型冬油菜生长点发育机理提供支撑。结果表明：白菜型油菜的BraSAUR基因共有142个,在10条染色体中不对称分布,主要分布在2、3号染色体上,基因长度较短,且大多数基因没有内含子。qRT-PCR结果表明,低温处理后,与CK相比,在两个品种的叶片中,Bra029452的表达量变化趋势不同,24 h时在L7中的表达量是X的24倍,在生长点中,Bra010501在L7中24 h时表达量达到CK的4倍,而在X中是先增加后降低的趋势。模拟干旱胁迫后,Bra029452在L7的叶片中的表达量均显著高于CK,在X中则是逐渐降低,在L7的生长点中1 h和24 h表达量均高于CK,但在X中则表现为先降低后升高。低温胁迫和干旱胁迫条件下,Bra029452基因表达量在强抗寒性油菜品种L7的叶片及生长点中均逐渐升高,可推断该基因同时参与白菜型冬油菜耐受低温、干旱的调控过程。     

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。     

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。     

向日葵3个谷胱甘肽-S-转移酶GST基因的克隆及表达模式分析

从向日葵转录组测序结果中获得了3个对盐胁迫有响应的谷胱甘肽-S-转移酶（GST,EC2.5.1.18）GST基因,构建了系统进化树并进行分析,得知这3个基因属于Tau型GST,并将它们命名为HaGSTU26（HanXRQChr04g0127901）、HaGSTU8（HanXRQChr06g0177581）、HaGSTU27（HanXRQChr10g0316331）,然后以向日葵Sk02R为试验材料,克隆这3个基因,并进行了不同组织和不同胁迫条件下的表达分析。序列分析表明：HaGSTU26基因组为1674bp,CDS（编码蛋白序列）长666bp,编码221个氨基酸,由2个外显子和1个内含子组成;HaGSTU8基因组为2271bp,CDS长654bp,编码217个氨基酸,由2个外显子和1个内含子组成; HaGSTU27基因组为663bp,CDS长663bp,编码220个氨基酸,由1个外显子组成。实时荧光定量PCR分析表明：向日葵HaGSTU26、HaGSTU8、HaGSTU27基因在不同组织（根、幼叶、成熟叶、茎、幼茎、苞叶）中表达量不同,其中,HaGSTU26基因和HaGSTU27基因在根中表达量最高,而HaGSTU8基因在苞叶中表达量最高,但这3个基因均在成熟茎中的表达量最低。在不同胁迫条件下,测定这3个基因在向日葵幼苗中的表达量,结果表明在盐及ABA胁迫下,基因表达量均随着处理时间的增加而呈现先增加后下降的趋势。其中,在盐胁迫下,HaGSTU26基因在12 h后相对表达量最高,HaGSTU8及HaGSTU27基因表达量在3h后相对表达量最高。在ABA胁迫后,HaGSTU26及HaGSTU27基因表达量在12 h后相对表达量最高,HaGSTU8在24 h后相对表达量最高。在冷胁迫下,HaGSTU26和HaGSTU27基因上调表达,它们分别在3、24 h后相对表达量最高,HaGSTU8基因下调表达,其相对表达量随处理时间的延长呈现逐渐减少的趋势。在热胁迫条件下,这3个基因的相对表达量随着胁迫时间的延长而增加,均在24 h后表达量最高。以上结果说明这3个基因对不同非生物胁迫（盐、ABA、冷、热胁迫）均有响应。     

苹果MdGH3-2/12在盐胁迫下的功能分析

于2020年5月—2021年2月以苹果野生型WT植株和MdGH3-2/12的RNA干扰转基因植株（RNAi）为材料，采用水培试验方法测定了盐胁迫条件下苹果植株的表型、生长量、光合参数、叶绿素含量、抗氧化酶及活性氧（ROS）含量变化，初步解析了MdGH3-2/12在苹果响应盐胁迫中的功能。研究结果表明：（1）与WT植株相比，RNAi植株叶表面出现更多的褐斑和坏死，且根系活力显著降低，其3个株系（GR-1,GR-9,GR-10）根系活力分别为WT植株的93.8%、77.5%和90.0%；植株的生长和光合作用受到更大影响，P_n、C_i和G_s都显著降低，其中P_n值分别下降至WT植株的62.9%、77.41%和83.6%。最大光化学效率（F_v/F_m）也显著下降，3个株系分别为WT植株的97.0%、96.4%和97.5%；叶绿素含量分别显著下降至WT植株的90.7%、86.2%和81.9%，MDA含量分别为WT植株的1.15、1.25倍和1.16倍；REL含量分别为WT植株的1.23、1.39倍和1.48倍；叶片气孔开度在盐胁迫下也分别缩小了35.0%、39.0%、55.0%和46.2%。（2）与WT植株相比，RNAi植株株系体内Pro含量显著增加，3个株系分别为WT植株的7.1、10.3倍和6.3倍；叶片中ROS积累显著高于WT植株，H₂O₂含量分别为WT植株的1.6、1.8倍和1.8倍，而抗氧化酶活性显著下降，各材料（WT,GR-1,GR-9,GR-10）盐胁迫组的SOD酶活性分别为对照组的2.4、1.2、1.7倍和1.9倍，这表明与WT植株相比，盐胁迫下RNAi植株各株系不能有效清除ROS，对植株造成了更大伤害。（3）盐胁迫后各材料植株的Na⁺/K⁺比值分别为0.497、0.558、0.525和0.577，RNAi植株株系显著高于WT植株，但是相关离子转运基因的表达量显著低于WT植株，致使Na⁺不能及时外排，植株受到钠离子毒害，从而导致其耐盐性降低。以上结果表明，MdGH3-2/12在苹果植株应对盐胁迫方面发挥着重要的作用，将MdGH3-2/12[STBZ]基因干扰后苹果植株在盐胁迫下受到的伤害更大，植株对盐胁迫的抗性下降。     

两个转W23基因小麦株系的抗旱性鉴定

以两个T₅代转W23基因小麦株系G19-X59、G19-X61为材料,在水培条件下采用PEG-6000人工模拟干旱胁迫措施对转基因小麦株系的根系发育特点、抗旱相关的光合生理等指标进行了测定。结果表明,干旱胁迫条件下各参试小麦的光合速率(Pn)和蒸腾速率(Tr)均下降,转基因株系降幅较小,且其Pn和Tr值极显著高于受体品种;在正常供水和干旱胁迫两种水分处理下,转基因株系均比受体品种具有较发达的根系,其根总长、根总表面积、根总体积等参数极显著高于受体品种。这一结果说明,转W23基因小麦株系G19-X59、G19-X61在苗期依靠发达的根系维持较强的光合作用,提高其应对干旱胁迫的能力。     

干旱胁迫对赤芍生理特性影响及相关基因的克隆表达

为探究不同干旱程度对赤芍生长发育的影响及其抗旱机制,采用盆栽控水法,设置对照组、轻度干旱和重度干旱3个处理,测定赤芍叶片相对含水量、可溶性糖、可溶性蛋白、脱落酸、叶绿素、类胡萝卜素、类黄酮、丙二醛、游离脯氨酸含量、光合参数及超氧化物酶活性等生理指标,评价赤芍抗旱性;采用RT-PCR结合RACE技术克隆抗旱相关基因（DREB、Psb27、PsaK、MDHAR、SOD和GR）,探究不同干旱程度对赤芍相关基因表达水平的影响。结果表明,重度干旱胁迫下,赤芍可溶性糖、可溶性蛋白、类胡萝卜素、类黄酮、脯氨酸、丙二醛、抗坏血酸及谷胱甘肽的含量均呈上升趋势,如在重度干旱和轻度干旱24 d后,赤芍叶片可溶性糖含量分别为对照的2.50倍和1.66倍,重度干旱胁迫第16天时,可溶性蛋白含量为对照组的2.43倍,类胡萝卜素与类黄酮的含量分别约为对照组的1.8倍和1.4倍,约为轻度干旱的1.4倍和1.2倍;而叶片相对含水量、蒸腾速率、气孔导度、净光合速率与叶绿素含量均降低,如重度干旱胁迫下赤芍叶片含水量不足20%,而轻度干旱和对照组叶片含水量均高于55%,叶绿素含量在第16天降到最低值,约为对照组的26%,为轻度干旱的33%;干旱胁迫下,光合作用相关基因Psb27、PsaK的表达量上调,且重度干旱上调水平更高;多个抗氧化基因的表达量（SOD、GR）、ABA含量以及胞间CO₂浓度呈先升后降趋势,如SOD在重度干旱第16天表达水平达到峰值,分别为轻度干旱与对照组的1.58倍和1.86倍,而后剧烈下降,到第24天时表达水平降到对照组的1.43倍,其中MDHAR的表达量呈下降趋势,重度干旱第24天时表达水平降到对照组的5.53倍。     

辣椒PP2C家族基因鉴定与响应低温胁迫的表达分析

对辣椒PP2C基因家族成员进行全基因组鉴定和特征分析,并研究CaPP2Cs基因在低温胁迫下的表达模式,结果表明：在辣椒参考基因组共系统鉴定出99个CaPP2Cs基因家族成员,并不均匀地分布在12条染色体上,CaPP2Cs基因编码的氨基酸数介于99~1 572 aa,分子量介于10.94~178.40 Da,等电点介于4.08~9.51。系统进化树分析可将辣椒和拟南芥的PP2C基因分为13个亚族,每个亚族均含有辣椒PP2C基因,基因结构分析表明CaPP2Cs基因的外显子数量介于1~18;在CaPP2Cs基因的启动子中发现许多与光、低温和激素响应有关的顺式作用元件。转录组数据分析表明,CaPP2C31基因在叶片中的表达量显著高于其他基因,CaPP2C16则特异性地表达于授粉后50 d的胎座中。低温处理24 h时,CaPP2C97和CaPP2C3在叶片中呈现显著的下调表达;低温处理1 h时,CaPP2C36、CaPP2C5、CaPP2C53、CaPP2C43、CaPP2C99等基因在根中明显上调,CaPP2C19、CaPP2C23、CaPP2C44、CaPP2C95则显著下调,表明CaPP2Cs可能在辣椒响应低温胁迫中发挥作用。     

干旱胁迫下不同中间砧对苹果苗生理特性及MdPIP1-1基因表达的影响

为探讨不同中间砧对‘天红2号’红富士苹果苗抗旱性的影响，以2 a生‘天红2号/黄6/平邑甜茶’、‘天红2号/SH40/平邑甜茶’、‘天红2号/GM256/平邑甜茶’盆栽苗为试材，研究了干旱胁迫下不同中间砧对苹果苗旱害指数、相关生理指标及水通道蛋白基因MdPIP1-1相对表达量的影响。结果表明，随着干旱胁迫时间延长，土壤含水量降低，3种中间砧苗旱害指数、O^-₂产生速率升高，F_v/F_m值降低，SOD、POD活性呈现先升高再降低的趋势，但3个砧穗组合各指标的变化幅度不同。干旱9 d时3个砧穗组合以‘天红2号/GM256/平邑甜茶’的旱害指数最高，为91.67%，其F_v/F_m值与正常供水相比显著下降11.47%；‘天红2号/黄6/平邑甜茶’旱害指数仅为29.55%，F_v/F_m值下降幅度最小。干旱胁迫期间，3种中间砧苗的MdPIP1-1基因表达量均呈现先升高再降低的趋势，黄6中间砧苗的表达量峰值出现在第7 d，GM256和SH40中间砧苗出现在第5 d。综合认为，3个砧穗组合抗旱性强弱依次为‘天红2号/黄6/平邑甜茶’＞‘天红2号/SH40/平邑甜茶’＞‘天红2号/ GM256/平邑甜茶’。     

Tumours of Begonia and some other ornamentals,induced by Corynebacterium fascians

In 1975 many tumours were observed in plants ofBegonia Schwabenland grown in Aalsmeer. Submersion of the roots ofNicotiana megalosiphon seedlings in a homogenate of tumorous tissue, induced tumours after two weeks. Short periods of submergence yielded results similar to those obtained after longer periods. Tumour homogenates lost their infectivity after ten min at 50°C. Aphids transmitted the infectious agent.Treatment with propylene oxide did not inhibit infectivity completely. Filtration through a 450 nm filter removed the infectious agent.Tobacco tumor virus or a viroid could not be isolated. Cultures ofCorynebacterium fascians, isolated from tumours ofN. megalosiphon were highly infectious and induced tumours in healthyN. megalosiphon andBegonia. Tumorous tissue homogenates ofPelargonium zonale, Dahlia sp.,Gladiolus sp., andLilium sp. also caused tumours inN. megalosiphon, from whichC. fascians was isolated. It was not possible to produce tumours inN. megalosiphon with homogenates from roses with symptoms of bud proliferation.Samenvatting In 1975 werden vele tumoren waargenomen inBegonia Schwabenland op Aalsmeerse bedrijven (Fig. 1). De infectiositeit van tumorweefsel kon goed en snel worden vastgesteld door de wortels van zaailingen vanNicotiana megalosiphon in een homogenaat van tumorweefsel te dompelen. Tumoren ontstonden na twee weken, de eindbeoordeling geschiedde na een maand (Fig. 2). Ook verschillende andereNicotiana spp.,Melilotus officinalis (Fig. 3) enPisum odoratum (Fig. 4) werden aangetast.Bij de infectiositeitstoets gaven zeer korte dompeltijden even goede resultaten als langere (Tabel 1). Infectieus sap verloor zijn infectievermogen na 10 min verhitting bij 50°C. Bladluizen brachten de smetstof over. Propyleenoxide verminderde de infectiositeit wel, doch onderdrukte deze niet totaal. Bij filtratie door een 450 nm filter bleef het infectieuse agens op het filter achter. Het tumor-inducerende agens was ook aanwezig in die delen van planten met tumoren welke gezond leken en het ging voor een gering deel over met zaad (Tabel 2).Uit tumoren konden wij geen tabakstumorvirus of een viroïde isoleren. Culturen vanCorynebacterium fascians, geïsoleerd uit tumoren vanN. megalosiphon bleken zeer infectieus en veroorzaakten tumoren inN. megalosiphon enBegonia. Homogenaten van tumorweefsel vanPelargonium zonale, dahlia (Fig. 5), gladiool (Fig. 6) enLilium Mid Century Hybrid Enchantment (Fig. 7) veroorzaakten ook tumoren opN. megalosiphon, waaruitC. fascians werd geïsoleerd. Met sap van kroeskopzieke rozen konden wijN. megalosiphon niet besmetten.     

Survey of Bacterial and Fungal Seedborne Diseases in Imported and Domestic Potato Seed Tubers

Potato seed tubers are imported to Israel from northern Europe and planted in spring; tubers harvested early from the spring crop are used as seed for the autumn crop. Although only seed lots registered as certified are imported, a previous survey (1984–1994) indicated that most imported lots were affected by latent or active infections caused byErwinia carotovora,Streptomyces scabies, Rhizoctonia solani, Fusarium spp. andSpongospora subterranae. The survey was extended until 1998, and included additional pathogens:Ralstonia solanacearum,Helminthosporium solani, Colletotrichum coccodes andVerticillium dahliae. Most of these pathogens were also monitored in domestic seed tubers, and are reported for the first time. Brown rot was not observed in any of the imported lots. Blackleg and soft rot caused byErwinia spp. were detected in most of the imported lots; however, less than 7% of the lots were contaminated at high levels, while approximately 65% were contaminated at moderate levels. Common scab was detected in most of the imported lots; 51% of the imported lots were contaminated at moderate or high levels, whereas only 6.5% of the domestic seed lots were contaminated at these levels. Black scurf was detected in most of the imported lots; on average, 47.3%, 44.2% and 1.4% of the lots were contaminated at low, moderate and high levels, respectively, and only 7.1% were disease-free. In contrast, most of the domestic lots were either disease-free (45.4%) or had a low disease incidence (37.3%). Only 16.7% of the lots were moderately infected and 0.2% were highly contaminated. Silver scurf was observed in most of the imported lots during all years of the survey, with no differences among the producing countries; on average, 22.7%, 66.1% and 7.5% of the lots were contaminated at low, moderate and high levels, respectively, and only 3.7% were disease-free. Most of the domestic lots (76%) were disease-free and only 6.6% were infected at moderate or high levels. Black dot was observed in a considerable portion of the shipments from Holland during all years of the survey, particularly in 1998, when 34% of the lots were infected. The shipments from France and Germany were infected at low levels, except in 1998, when 19% and 11% of the lots, respectively, arrived infected. In shipments from Scotland and Ireland low incidences of the disease were observed in 1994 and 1995. In the domestic lots, black dot incidence was low (<2.4%) except in 1996, when 11% of the lots were infected.V. dahliae was monitored only in domestic seed tubers. The incidence of disease-free lots was 56–64%, whereas in 20–30% of the lots the level of infection was <5%, and in 6–16% of the lots the level was >5%. The survey findings demonstrate transmission of seedborne pathogens; most of these pathogens can become established in the soil and eventually cause severe outbreaks of disease in potatoes grown in Israel. http://www.phytoparasitica.org posting May 16, 1999.     

Epidemiology and simulation of population development of Sitobion avenae in winter wheat

The epidemiology ofSitobion avenae and its natural enemies in winter wheat was studied in 1975, 1976 and 1977. Immigration was important until the end of flowering. The alate immigrants had apterous offspring. These became the driving force in population growth. Their offspring were mostly alatae which usually left the field. A model of the epidemic was developed. Quantitative relations between the aphids and their environment were obtained from literature or established in laboratory trials. The model simulated population development and population composition from the beginning of June till the population peak at the end of June or early in July. Because quantitative data on relations between aphids and their natural enemies and pathogens are scarce, and since the knowledge on wing formation is still limited, the population collapse could not be predicted. In the future, prognosis over a period of three weeks seems possible.Samenvatting De toenemende betekenis van graanbladluizen (vooralSitobion avenae) gepaard gaande met een sterke toename van het gebruik van insecticiden op granen maakte verbetering van de prognose over het schadelijk optreden wenselijk. Door gedetailleerde tellingen in het veld (Fig. 1–7) werden gegevens verkregen over het verloop van de epidemie en het optreden van natuurlijke vijanden in 1975, 1976 en 1977.Een immigratieperiode tot in de bloei kon worden vastgesteld. Daarna lijkt de aantrekkelijkheid van het gewas voor alate luizen te verminderen. De alate immigranten krijgen aptere nakomelingen. Deze vormen de stuwende kracht van de populatiegroei. De nakomelingen van apteren zijn merendeels alaat. Zij verlaten het gewas.Een model van de populatieontwikkeling gedurende de epidemie werd opgesteld. De relatiediagrammen Fig. 9 en 10 laten groei en ontwikkeling vanS. avenae en een predator (Syrphus corollae) zien. Kwantificering van de betrekkingen werd mogelijk door literatuurgegevens en laboratoriumexperimenten.Met het model kon de populatieontwikkeling vanS. avenae vanaf begin juni tot aan de populatiepiek in 1975, 1976 en 1977 vrij goed worden gesimuleerd (Fig. 12). Ook de populatieopbouw kon worden gesimuleerd (Fig. 14). De teruggang van de populatiedichtheid blijkt moeilijker te voorspellen door het ontbreken van gegevens over natuurlijke vijanden.Het lijkt waarschijnlijk dat in de toekomst met het model een prognose over de piek van de bladluispopulatie circa 3 weken tevoren mogelijk zal zijn.     

Recent observations on leafy gall in Liliaceae and some other families

Corynebacterium fascians, which causes leafy gall, has been shown to be responsible for the unusual symptoms recently found in lilies. The symptoms are described and the different bacterial isolates compared by biochemical, serological and pathogenicity experiments. No evidence could be found for the existence of specialized strains even though considerable variation in virulence could be demonstrated. Although the role of variation in susceptibility of the different lily cultivars should not be underestimated, it would appear that high inoculum levels ofC. fascians in the soil may be largely responsible for these outbreaks.Also included in this study are the results of biochemical, serological and pathogenicity experiments ofC. fascians which has been isolated fromKalanchoe, Euphorbia, Brodiaea, Hebe andVerbascum.Samenvatting Reeds een aantal jaren was in lelies een woekering in stengelbollen bekend, die echter sporadisch voorkwam en economisch van weinig betekenis was. Tijdens de zomer van 1977 echter trad de ziekte op één enkel perceel zo ernstig op dat zeer aanzienlijke oogstderving het gevolg was. Behalve de stengelbol kan ook de hoofdbol misvormd zijn. Bovendien kunnen misvormde stengelbollen lange spruiten vormen, zgn. sabeltandvorming. Soms is het wortelstelsel gereduceerd; in de kas zijn verdikte stengels waargenomen. De ziekte staat nu bekend onder de naam woekerziekte.Door de PD te Wageningen en het LBO te Lisse werd de bacterieCorynebacterium fascians (Tilford) Dows. uit het zieke materiaal geïsoleerd. In gezamenlijk onderzoek werd nagegaan of de bacterie de veroorzaker is van bovengenoemde symptomen en of hier sprake is van een op lelie gespecialiseerde stam.Biochemische en serologische vergelijkingen tussen isolaten vanC. fascians uit lelie,Kalanchoe, Euphorbia, Brodiaeea, Hebe enVerbascum laten zien dat er zeer weinig verschillen bestaan. Alle isolaten reageren op een standaard antiserum met titers variërend tussen 1: 640–1: 5120.Een oriënterende inoculatieproef werd uitgevoerd op het LBO in 1977. Daarna werden op de PD 10 van de 25 isolaten uit lelie en 8 uit andere waardplanten getoetst op lelie, chrysant en lathyrus. Uit de inoculatieproeven blijkt, dat de bacterie de veroorzaker is van de waargenomen symptomen. Tussen de isolaten blijkt een groot verschil in pathogeniteit te bestaan (12–100% aantasting), zonder dat dit aan herkomst gebonden is. Het voorkomen van op lelie of andere waardplanten gespecialiseerde stammen is daarom niet aan te nemen.In hoeverre de hevige aantasting een gevolg is van een opbouw van de bacterie-populatie door een te nauwe vruchtwisseling en mechanische rooimethoden moet worden nagegaan.     

Spatial pattern of Cercospora leaf spot of sugar beet in fields in long- and recently-established areas

Spatial disease pattern of Cercospora beticola was characterised during natural epidemics of Cercospora leaf spot (CLS) in sugar beet. We applied linear regression and geostatistical analyses to characterise CLS spatial patterns in three field trials, in long-established and recently-established CLS-areas, during two consecutive years. Linear regression showed a positive influence of average disease severity of within-row neighbouring plants (0.38 < < 0.88). Semi-variograms modelled the spatial dependence of disease severity for two directions per week in both years. Disease severity displayed strong spatial dependence over time. The within-row spatial dependence was the largest, but across-row dependence was irregular and weaker. Both long- and recently established areas showed strong spatial dependence of disease severity within row, decrease in variability between years and within the second trial year and a relation between and the relative nugget. Observed differences were more field than area specific. These spatial and temporal analyses indicated that disease severities of adjacent plants were dependent; hence, we concluded that C. beticola is dispersed mainly over short distances from plant to plant.     

Identification and pathogenicity of Pythium species causing damping-off of kidney bean

Five Pythium species (Pythium irregulare, P. mamillatum, P. myriotylum, P. spinosum and P. ultimum var. ultimum) were isolated from the hypocotyls and roots of kidney bean plants with damping-off from a commercial field and from experimental plots that have undergone either continuous cropping with kidney bean or rotational cropping with arable crops. In inoculation tests, all five Pythium species were pathogenic to kidney bean. This is the first report of damping-off of kidney bean caused by Pythium species; we named this disease damping-off of kidney bean. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accession numbers AB291811, AB291944 and AB291945.     

Pathogenic fungi involved in root rot of peas in the Netherlands and their physiological specialization

Research on root rot pathogens of peas in the Netherlands has confirmed the prevalence ofFusarium solani, F. oxysporum, Pythium spp.,Mycosphaerella pinodes andPhoma medicaginis var.pinodella. Aphanomyces euteiches andThielaviopsis basicola were identified for the first time as pea pathogens in the Netherlands. Other pathogens such asRhizoctonia solani andCylindrocarpon destructans were also found on diseased parts of roots. F. solani existed in different degrees of pathogenicity, and was sometimes highly specific to pea, dwarf bean of field bean, depending on the cropping history of the field.A. euteiches was specific to peas, whereasT. basicola showed some degree of physiological specialization.     

Detection of the phytotoxin coronatine by ELISA and localization in infected plant tissue

Several pathovars of Pseudomonas syringae produce the phytotoxin coronatine (COR)in vitro, and the availability of purified toxin has facilitated development of immunological detection methods. A modified, indirect competitive ELISA using the COR-specific monoclonal antibody 11B8 was developed to detect COR in various host plants infected by P. syringae. The estimated detection limit for COR was 50 pg per well, and COR could be reliably quantified from 5 to 40 ng ml⁻¹. The subcellular localization of COR within infected tomato tissue was investigated using the COR-specific antibody MAb 8H3G2. Immunofluorescence microscopy and immunogold labelling showed that COR was present inside tomato cells and was associated with chloroplasts and particles of proteinase inhibitor I. Localization studies indicated that COR is mobile in infected plant tissue and can be detected in healthy tissue adjacent to the bacterial lesions.     

Biocontrol agents in combination with Moringa oleifera extract for integrated control of Sclerotium-caused cowpea damping-off and stem rot

Damping-off and stem rot disease-causing Sclerotium rolfsii has been reported as a destructive soil-borne pathogen of numerous crops, especially in the tropics and subtropics. Trials were conducted to test the efficacy of biocontrol agents alone or combined with Moringa oleifera leaf extracts for the control of the disease. In the laboratory, PDA was amended with Moringa leaf extract, and mycelial growth of S. rolfsii was measured. In the greenhouse and field, Trichoderma Kd 63, Trichoderma IITA 508 and Bacillus subtilis were evaluated as seed treatments, soil drench or sprinkle, separately or combined with Moringa leaf extracts. Percentage disease incidence, severity and control were recorded. In the laboratory, the higher the extract concentration the less the mycelial growth and no mycelial growth occurred on extract at 15 or 20 g leaves 10 ml⁻¹ water. In the greenhouse, the highest disease control was observed at a Moringa extract concentration of 15 kg leaves 10 l⁻¹ water (w/v). Seed treatments using Trichoderma Kd 63, and soil sprinkle using Trichoderma IITA 508 had a significantly (P = 0.05) higher effect on a disease incidence than Bacillus. Disease severity followed the same pattern. Moringa seed treatment combined with Trichoderma soil sprinkle resulted in significantly more than 94% and 70% disease control in the greenhouse and field, respectively, with significant yield increase in the field. This is the first report of Moringa leaf extract combined with Trichoderma as an integrated control for Sclerotium damping-off and stem rot of cowpea in the field.     

Bacterial induction of the accumulation of phaseollin,pisatin and rishitin and their antibacterial activity

Bean hypocotyls, pea pods and tomato fruits were tested for phaseollin, pisatin and rishitin production when challenged with the phytopathogenic bacteriaErwinia carotovora, Pseudomonas phaseolicola, P. pisi andP. solanacearum, and their isolated extracellular polysaccharides. All bacteria induced phytoalexin accumulation, whereas only phaseollin and pisatin, but not rishitin, were elicited by EPS. The inhibitory effect of these three phytoalexins on bacterial growth was studied in liquid medium; whereas phaseollin and pisatin strongly inhibited growth, only a slight inhibitory effect resulted from the presence of rishitin in the medium.Samenvatting Bonehypocotylen, erwtepeulen en tomatevruchten werden onderzocht op hun vermogen tot vorming van respectievelijk faseolline, pisatine en rishitine, na inoculatie met de fytopathogene bacteriënErwinia carotovora, Pseudomonas phaseolicola, P. pisi enP. solanacearum en na behandeling met oplossingen van hun extracellulaire polysacchariden (EPS). Alle bacteriesoorten induceerden fytoalexinevorming, terwijl hun EPS wel faseolline- en pisatine-, maar geen rishitinevorming induceerden. Faseolline en pisatine remden de groei van de bacteriën in vloeibaar medium sterk; rishitine daarentegen had slechts een geringe groeiremming tengevolge.