多肉植物翡翠景天黑腐病病原菌

福建漳州是景天科Crassulaceae多肉植物翡翠景天Sedum morganianum主产地。近年来,该植物黑腐病发病严重,为鉴定其病原菌,从福建漳州3个翡翠景天种植基地采集典型病株并分离病原菌,基于形态学观察及rDNA-ITS、actin、EF-1α基因序列分析和致病性测定,界定该病原菌为山扁豆生棒孢Corynespora cassiicola。该菌引起多肉植物翡翠景天黑腐病为世界首次报道。     

广东省柑橘炭疽病病原菌的形态与分子鉴定

炭疽病是柑橘的主要真菌性病害之一。2007年春,广东省德庆县名优柑橘品种贡柑炭疽病暴发流行。为了明确该县及广东省其他地区柑橘炭疽病菌的种类,为防治提供依据,对采集自广东省6个地区柑橘属10个栽培品种上的炭疽病样本进行病原菌分离,共获得柑橘炭疽病菌单孢菌株75株,对其中10株代表性的菌株进行了种类鉴定。通过培养性状和形态学特征观测、核糖体DNA(rDNA)内转录间区(ITS)序列分析、ITS区特异性引物PCR检测和系统发育关系比较等方面的研究,结果表明:10个柑橘炭疽病菌菌株均为盘长孢状刺盘孢Colletotrichum gloeosporioides,未发现国际上其他国家报道的严重危害柑橘花器和幼果部位的柑橘花后落果病病原菌——尖刺盘孢C.acutatum。     

bZIP转录因子CfHac1参与调控果生刺盘孢菌的生长发育和致病力

油茶炭疽病是油茶Camellia oleifera上最重要病害之一,引起该病害的主要致病菌为果生刺盘孢菌Colletotrichum fructicola。本研究以果生刺盘孢菌bZIP类转录因子CfHac1为研究对象,研究其在果生刺盘孢菌的营养生长、产孢量、附着胞形成、致病力及耐受性等方面的生物学功能,为油茶炭疽病的防控提供理论依据。研究结果表明,果生刺盘孢菌中具有一个与灰色大角间座壳（稻瘟菌）bZIP转录因子MoHac1直系同源的基因,命名为CfHAC1。该基因全长1 627bp,编码526个氨基酸,该蛋白含有一个碱性亮氨酸链（bZIP）结构域和3个未知功能结构域。CfHAC1基因敲除突变体的菌丝生长速度显著变慢,分生孢子产量显著减少且不能正常形成附着胞,并对山梨糖醇和KCl渗透压胁迫敏感性增加;致病力测试结果表明,果生刺盘孢菌基因敲除突变体ΔCfhac1对油茶的致病力显著下降。转录因子CfHac1参与调控果生刺盘孢菌的生长、产孢、附着胞的形成、致病力以及响应外界渗透压胁迫过程。     

组蛋白去乙酰化酶CfSnt2调控果生刺盘孢生长发育和致病力

炭疽病是油茶Camellia oleifera的重要病害,该病害的优势致病菌是刺盘孢属Colletotrichum的果生刺盘孢C. fructicola,在全国的油茶产区普遍发生。我们前期发现组蛋白乙酰转移酶CfGcn5调控油茶果生刺盘孢的生长发育和致病过程,但组蛋白去乙酰化酶在该病菌中的生物学功能尚不清楚。本研究以组蛋白去乙酰化酶CfSnt2为研究对象,利用反向遗传学的方法,通过对野生型、CfSNT2基因敲除突变体及互补菌株的生物学表型进行比较分析,发现CfSNT2基因敲除突变体的菌丝生长速率明显减缓、分生孢子的产量显著减少、附着胞形成率降低、对细胞壁胁迫剂的响应异常,同时对油茶致病力显著减弱。以上现象说明CfSnt2调控果生刺盘孢的生长、产孢、附着胞的形成、对细胞壁完整性胁迫剂的耐受性及致病力。     

抗辣椒炭疽病菌盘长孢状刺盘孢的木霉菌的筛选

本试验通过对峙培养法筛选出28株具有强拮抗能力和15株中等拮抗能力的木霉菌。使用玻璃纸培养法进一步复筛出6株木霉菌(104、393、424、402、451和123T),其非挥发性次生代谢产物对盘长孢状刺盘孢的抑制率高达92%以上。通过平板对扣培养法发现木霉菌424(Trichoderma atroviride)以及123T (T. paratroviride)的挥发性次生代谢产物对盘长孢状刺盘孢也具有抑制作用,抑制率分别为57.12%和60.43%。试验还对9个辣椒品种进行了盘长孢状刺盘孢的果实致病性评价,发现红尖椒属于高度易感病品种。在对该品种进行针尖刺伤法处理后,喷施木霉菌424和123T的分生孢子悬浮液后可以提高其对病菌的抗性。此外,试验还发现上述2株木霉菌的孢子悬浮液对辣椒种子萌发及幼苗生长具有促进作用,其将辣椒种子的发芽率由38.9%提高至70.0%。同时,辣椒幼苗经木霉菌分生孢子处理后,株高、根长、茎粗、鲜重和干重分别增加了52.5%、96.0%、104.0%、372.0%和224.0%。综上所述,本试验筛选的木霉菌424和123T能够充分发挥其生防能力,为进一步开发辣椒炭...     

‘海螺’望春花内生真菌EF-WCH-51鉴定及其抑制葫芦科刺盘孢活性成分分析

【背景】药用植物内生真菌产生的次生代谢产物具有多种抗菌活性,因而具有生防菌开发潜力。【目的】鉴定一株对葫芦科刺盘孢(Colletotrichumorbiculare)有抑菌活性的‘海螺’望春花内生真菌EF-WCH-51,并确定其主要抑菌活性成分。【方法】根据形态特征并结合分子序列鉴定EF-WCH-51菌株,利用生长速率法确定EF-WCH-51抑菌活性,并采用半制备型HPLC、HPLC-MS/MS对其活性成分进行分离鉴定。【结果】供试菌株EF-WCH-51为丝枝蜡蚧菌(Lecanicillium aphanocladii),其发酵液粗提物对葫芦科刺盘孢菌丝生长具有较强抑制作用,EC50为0.086 2 mg/mL;抑菌活性组分A (0.085 0 mg/mL)对葫芦科刺盘孢抑制率达到76.08%,通过查询MS、MS/MS数据库以及与相关文献对比,确定该化合物为卵孢菌素(oosporein)。【结论】本研究发现丝枝蜡蚧菌对葫芦科刺盘孢具有较好的抑菌活性,其主要抑菌活性成分为卵孢菌素,具有作为瓜类炭疽病生防菌株的潜力。     

莴笋炭疽病病原鉴定

【背景】2018年7-8月,甘肃省兰州市永登县武胜驿镇种植的莴笋大面积发生炭疽病,约40%的地块发病,绝收面积达10%。【目的】明确莴笋炭疽病的病原。【方法】采用组织分离法进行病原菌分离;通过Koch’s法则明确分出病菌的致病性;采用形态学和分子生物学方法对病原菌进行种类鉴定。【结果】分离得到形态特征一致的真菌菌株3株。在PDA平板上20°C培养7 d,病菌分生孢子无色,单隔,梭形,(10.44-19.40)μm×(2.61-4.48)μm。代表性菌株Lett-11接种莴笋离体叶片可引起与自然发病相似的症状。BLASTn分析结果显示,菌株Lett-11 (GenBank登录号MK252097)的r DNA-ITS序列与莴苣盘二孢菌Marssonina panattoniana strain CBS 163.25 (GenBank登录号MH854831.1)的序列相似性达99%。【结论】引起莴笋炭疽病的病原菌被鉴定为莴苣盘二孢菌Mar. panattoniana [Synonymy:Microdochium panattonianum],这是莴苣盘二孢菌引起莴笋炭疽病在甘肃的首次报道。     

对进境高粱种子中高粱炭疽病菌疏纹刺盘孢的检疫

从一批进境的美国高粱种子样品中分离得到一株与疏纹刺盘孢Colletotrichum sublineola形态相似的菌株6222-1。经形态学观察发现该菌产生大量镰状或梭形分生孢子,在菌丝末端形成膨大的附着胞。经多位点基因(ITS、GAPDH、CHS-1、HIS3、ACT、TUB2)系统发育分析,发现该菌株与C.sublineola模式菌株聚集在同一个分支上,支持率达到100%。经病原菌接种高粱幼苗叶部,引起典型的炭疽病症状。根据上述研究结果,确认该菌株为疏纹刺盘孢。这是我国首次截获该危险性病原真菌。     

健康与患病刺梨植株可培养叶际真菌菌群差异比较

本研究比较了健康与患叶斑病刺梨植株叶际真菌群落特征差异,以期探索病原菌的潜在来源,为人工构建具拮抗功能群落和刺梨叶斑病的生物防控提供参考。通过可培养方法对不同健康状况的刺梨叶际真菌进行分离培养,结合形态学和分子系统学对菌株进行综合鉴定;利用FUNGuild平台对真菌进行功能注释;结合根际、根部真菌作拆分网络分析探索病原菌的潜在来源。本研究结果表明：1）刺梨叶际真菌具有丰富的多样性。从刺梨叶际8个样品中共分离到真菌266株,其隶属于3门、6纲、13目、30科、46属中的61个种。其中,健康植株叶际内生真菌（LHE）包括8属10种27株,附生真菌（LHS）包括33属37种77株。患病叶际内生真菌（LDE）分离到7属10种38株;附生真菌（LDS）分离到31属35种124株。2）不同样品的真菌优势属和特有类群有差异。不同健康状况下叶际附生真菌的优势属均为拟盘多毛孢属Pestalotiopsis,但二者的相对多度存在差异,LHS为11.49%,LDS为32.26%;内生真菌优势属二者均为链格孢属Alternaria,但相对多度各异,LHE为33.33%,LDE为63.16%。其中,LHE特有类群为盘长孢状刺盘孢 Colletotrichum gloeosporioides和果生刺盘孢Colletotrichum fructicola等8种;LDE特有类群为茄链格孢 Alternaria solani和Didymella sinensis 等8种;LHS特有类群是草酸青霉Penicillium oxalicum和Peniophora crassitunicata等21种;LDS特有类群是尖孢镰刀菌Fusarium oxysporum、赭绿青霉Penicillium ochrochloron和易脆毛霉Mucor fragilis等19种。3）不同样品叶际真菌功能不同。经FUNGuild解析表明,LHS、LHE和LDE的叶际真菌功能群主要以腐生型为主,LDS则主要以植物病原菌群为主。本研究结果初步揭示刺梨植株健康与患叶斑病叶际间真菌多样性、群落组成及营养功能群存在差异,植株健康状况与其真菌群落特征密切相关;叶斑病病原菌主要源于刺梨叶际的附生微生物群。     

果生刺盘孢CfHAC1调控应答二硫苏糖醇胁迫的转录组分析

果生刺盘孢Colletotrichum fructicola是油茶炭疽病优势病原菌。前期研究发现bZIP转录因子CfHac1参与调控该菌的生长发育和致病性。为了揭示转录因子CfHac1调控果生刺盘孢响应内质网压力和致病机理,本研究测定了ΔCfhac1突变体对内质网压力胁迫剂的敏感性,发现突变体对二硫苏糖醇（dithiothreitol,DTT）的耐受性下降,说明CfHAC1基因可能参与调控果生刺盘孢响应内质网压力胁迫过程。进一步利用高通量RNA-seq技术对该病菌野生型菌株和CfHAC1敲除突变体菌株在DTT胁迫下的转录组进行了比较分析,结果表明差异表达基因共有2 680个,其中上调表达基因有1 181个,下调表达基因有1 499个。Gene Ontology 功能分析结果显示,差异表达基因主要参与催化活性、结合、代谢过程、细胞过程、细胞成分合成、生物过程调控和应激反应等生物学过程。KEGG功能富集分析表明,上调表达基因主要被富集到核糖体、真核细胞的核糖体生物合成、RNA转运和氰基氨基酸代谢通路中;下调表达基因显著富集在内质网蛋白质加工、N-聚糖生物合成、类固醇合成和蛋白质分泌等通路中。分析发现转录因子CfHac1调控内质网胁迫应答和致病相关基因的表达。本研究提供了在全基因组水平上对CfHAC1基因与内质网压力胁迫应答之间关联的新认识,为阐明果生刺盘孢响应内质网压力胁迫和致病机制奠定了基础。     

珍稀濒危植物细果秤锤树开花生物学特性和繁育系统

为了解珍稀濒危植物细果秤锤树（Sinojackia microcarpa）开花特征和有性繁殖,对其花部形态特征、开花动态、花粉胚珠比（P/O）、柱头可授性、花粉活力、套袋试验、访花昆虫及访花频率进行观测。结果表明：（1）细果秤锤树的杂交指数（OCI）为4,单花期5~7 d,种群花期可持续20 d左右。花粉与胚珠比为4 093.21±498.56。开花后第3天的花粉活力最高（76.21%）,而开花后第7天时花粉活力较低（18.37%）。细果秤锤树柱头最适可授期在开花后第2天。（2）套袋试验表明,细果秤锤树存在部分自交不亲和性,同时不存在无融合生殖,传粉昆虫是其完成生殖过程所必需的,且异株授粉能够提高其坐果率和结籽率。细果秤锤树的访花昆虫有3目5科7种,主要访花昆虫有黄胸木蜂（Xylocopa appendiculata）、熊蜂（Bombus sp.）、胡蜂（Vespa sp.）、中华蜜蜂（Apis cerana）、黑带食蚜蝇（Episyrphus balteatus）、其中熊蜂平均访花频率最高,达（8.67±0.21） 次·h^-1。对该物种开花生物学特征与繁育系统进行深入研究,有利于进一步探究其濒危机制,为后续珍稀濒危植物的研究提供理论依据和参考。     

Chemical composition of volatiles from the syconia of Ficus microcarpa and host recognition behavior of pollinating fig wasps

Aims Chemical communication plays a key role in host plant recognition of pollinators. There are two recognized types of chemical communication between syconia and their pollinating fig wasps: one is “generalization”, of which the wasps respond to the relative ratio of multiple compounds, and the other is “specialization”, of which the key signal is a single uncommon, possibly unique, compound. The aims of this study were to identify the chemical composition of volatiles from the syconia of Ficus microcarpa at different developmental phases, and to determine if the signaling between F. microcarpa and its pollinating fig wasp, Eupristina verticillata, is of generalized type, or of specialized type.Methods The volatiles from syconia of F. microcarpa were extracted using solid-phase micro extraction (SPME) at different developmental phases (pre-female, female (before and after pollination), interfloral, male and postfloral phases) and the chemical compounds were identified by gas chromatography mass spectrometry (GC-MS). We then tested the behavioral responses of E. verticillata to fresh syconia at different developmental phases using two-choice olfactometers.Important findings There were 21 volatile compounds identified from the syconia at different developmental phases, which were mainly fatty acid derivatives, terpenoids and aromatic compounds. The components of the volatiles apparently differed among the developmental stages. The contents of terpenoids declined, but the contents of fatty acid derivatives increased, from before the pollination to after the pollination. Especially, the characteristic compounds of 2-heptanone and 3-octanone before the pollination disappeared, D-limonene decreased after the pollination, but copanene, cyclohexane and 2-hexenal increased. The results of the two-choice olfactometer experiment showed that the pollinating fig wasps had higher selection ratio to chemicals found in the female phase syconia than those in other phases; whereas the volatile compounds from the male phase syconia had the function pushing the pollinating fig wasps to leave the natal syconia so that there existed the “push-pull” responses by fig wasps to volatiles released by their host syconia. We conclude that there are multiple chemical compounds playing the roles in host recognition of pollinating fig wasp E. verticillata. The mutualistic relationship between F. microcarpa and E. verticillata is maintained by the chemical communication of “generalization” strategy.     

基于叶绿素荧光成像及荧光参数分布特征的叶片光合异质性定量分析

植物叶片光合作用具有高度的空间异质性,叶绿素荧光成像技术为叶片光合异质性的研究提供了便利,但叶片光合异质性的定量分析并没有得到广泛应用。本文利用Imaging PAM叶绿素荧光成像系统,获得中亚热带地区越冬期小叶榕(Ficus microcarpa)阳生叶和阴生叶的叶绿素荧光参数图像,并利用仪器的分析软件对其进行分析,定量比较了阳生叶和阴生叶的光合异质性特征。研究发现：越冬期小叶榕阳生叶的光合异质性和光抑制程度明显高于阴生叶,变异系数可作为光合异质性的定量指标。低温强光导致阳生叶坏死率(P_LN)达4.30%,并有53.30%的区域处于严重光抑制(0v/F_m<0.627),但仍有42.27%的区域仅为轻度光抑制(0.627≤F_v/F_m<0.800)。而低温弱光并未造成阴生叶坏死和严重光抑制。通过对光系统Ⅱ(PSⅡ)的实际光合效率(Y(Ⅱ))、调节性能量耗散的量子产额(Y(NPQ))和非调节性能量耗散的量子产额(Y(NO))荧光参数异质性的定量分析表明,阳生叶...     

后河保护区柳杉林中黏菌群落组成

柳杉是中国特有树种,对于广泛分布于森林生态系统的黏菌而言,单纯的柳杉林是一种特殊的生境。为探索柳杉林中黏菌的物种多样性,本研究在武陵山区东北部的湖北省后河国家级自然保护区柳杉林样地中采集柳杉树皮和凋落物,在120个培养湿室中,发生黏菌的湿室68个,占56.67%。从两种基物上获得黏菌6目7科12属24种,优势种为混淆筛菌Cribraria confusa,相对丰度达到19.01%。柳杉林凋落物上发生的黏菌种类和数量高于柳杉树皮,凋落物和柳杉树皮上发生的黏菌共有种为4种,种群间的Sørensen相似性系数为Cs=0.333。在不同季节间,黏菌物种的群落组成与多样性也表现出差异,春季Shannon-Wiener多样性指数Hʹ=0.2987±0.1564,夏季Hʹ=0.1105±0.0949,秋季Hʹ=0.1968±0.0589,冬季Hʹ=0.2726±0.0875。黏菌物种数量发生最多的是春季和冬季,均有15种,仅在春季发生的黏菌物种为粗壮发菌Comatricha suksdorfii、鲜黄绒泡菌Physarum luteolum、暗红团网菌Arcyria denudata和极小无丝菌Licea minima,仅在冬季发生的黏菌物种为钙丝绒泡菌Physarum decipiens、小筛菌Cribraria microcarpa和暗褐无丝菌Licea belmontiana。典型对应分析（canonical correspondence analyses,CCA）表明,影响柳杉林中黏菌物种群落组成与多样性的主要环境因素是基物类型。     

两种生态型榕树的叶绿素含量、荧光特性和叶片气体交换日变化的比较研究

比较盆栽 生榕树和两栖型树的形态差异、叶片叶绿素含量、叶绿素荧光特性和气体交换的日变化。两栖型榕树具有较发达的气生根和水生不定根,叶片比陆生榕树宽,并有向中生性 倾向,陆生榕树的叶绿素含量比两栖榕树高,净光合速率略高于水培两栖型榕树,但明显高于土培两栖型榕树,蒸腾速率以水培两栖型树最高,陆生榕树次之,土培两栖型榕树最低,线性回归分析表明,三者的叶片气孔导度与净光合速率变化均呈正相关,气孔导度的变化     

木论喀斯特常绿落叶阔叶混交林群丛数量分类及稳定性

加深对喀斯特顶极群落植物组成、群落结构和群落分布的认识可以为该区域生物多样性保护和森林管理提供参考。本文基于广西木论喀斯特常绿落叶阔叶混交林25 ha森林动态监测样地地形、土壤和物种组成数据, 采用多元回归树对该群落进行群丛分类, 并分析各群丛多样性和稳定性。结果表明, 群落可分为6个群丛, 分别为群丛I: 长序厚壳桂+栀子皮+香叶树群丛(Ass. Cryptocarya microcarpa + Itoa orientalis + Lindera communis), 群丛II: 长序厚壳桂+灰岩棒柄花+罗伞群丛(Ass. Cryptocarya microcarpa + Cleidion bracteosum + Brassaiopsis glomerulata), 群丛III: 圆果化香树+密花树+齿叶黄皮群丛(Ass. Platycarya longipes + Rapanea neriifolia + Clausena dunniana), 群丛IV: 圆果化香树+滇丁香+齿叶黄皮群丛(Ass. Platycarya longipes + Luculia intermedia + Clausena dunniana), 群丛V: 长序厚壳桂+罗伞+伞花木群丛(Ass. Cryptocarya microcarpa + Brassaiopsis glomerulata + Eurycorymbus cavaleriei), 群丛VI: 小叶栾树+长管越南茜+圆果化香树群丛(Ass. Boniodendron minus + Rubovietnamia aristate + Platycarya longipes)。除群丛I外, 各群丛总体的多样性指数较高。Shannon-Wiener指数、Simpson指数以及Pielou均匀度指数表现出一致的变化趋势: 群丛VI > 群丛V > 群丛IV > 群丛III > 群丛II > 群丛I, 而丰富度则为群丛VI > 群丛IV > 群丛V > 群丛I > 群丛III > 群丛II, 物种多样性在中海拔群丛最高。中上坡部位群丛稳定性最高, 洼地群丛稳定性最低。海拔在群落结构及组成中起重要作用, 可能是影响群落分布的重要因素。     

Can Flowering Plants Enhance Numbers of Beneficial Arthropods in UK Apple and Pear Orchards?

A replicated experiment with fourteen flowering plant species was used to determine the effectiveness of flowering plants as attractants for beneficial arthropods in orchards. The greatest numbers of anthocorids were found on cornflower (Centaurea cyanus) and corn chamomile (Anthemis arvensis) and hymenopteran parasitoids on corn marigold (Chrysanthemum segetum) and corn chamomile. The impact on pest numbers of manipulating the orchard environment by the use of flowering plants was investigated in two ways. In an apple and a pear orchard, a proportion of the trees was undersown with a mixture of cornflower, corn marigold and corn chamomile. No significant differences were found in numbers of Panonychus ulmi (in apple) or Cacopsylla pyricola (in pear) on trees grown in the undersown areas compared with trees grown in conventional bare herbicide strips. When potted pear trees infested with pear psyllids were placed either into plots sown with the flower mixture or on bare earth, psyllid numbers declined rapidly on trees in both treatments. When beneficials were excluded from the potted control trees, numbers of psyllid larvae, but not eggs, declined more quickly on the trees surrounded by flowering plants. The importance of habitat diversity on biocontrol in orchards is discussed.     

Suppression of aphids on strawberry by augmentative releases of larvae of the lacewing Chrysoperla carnea (Stephens)

In laboratory feeding experiments, larvae of Chrysoperla carnea consumed up to 790 Chaetosiphon fragaefolii during their development. In gauzehouse experiments, releases of C. carnea larvae significantly reduced numbers of C. fragaefolii on potted strawberry plants. In open field experiments numbers of C. fragaefolii were significantly reduced at release rates of eight chrysopid larvae per plant, but a release experiment on larger strawberry plants grown under protection did not give a significant reduction in aphid numbers, even at a release rate of 25 larvae per plant.     

Survival of larvae of the ragwort flea beetle, Longitarsus flavicornis (Coleoptera: Chrysomelidae), in water-logged soil

Water-logged soil could be a major factor affecting populations of the ragwort biological control agent, Longitarsus flavicornis, in Australia. Survival of L. flavicornis larvae in potted ragwort plants after inundation with water for up to 264h (11days) was compared to larval survival in control plants not exposed to inundation. Probit analysis indicated that 35.6% mortality of the larval population occurred after 24h inundation and 50.4% after 72h. As high mortality was recorded at periods of inundation known to occur in the field, it is concluded that winter flooding is likely to be detrimental to populations of L. flavicornis.     

AM真菌摩西管柄囊霉对干旱胁迫下枳抗氧化酶基因表达的影响

测定分析了接种丛枝菌根（AM）真菌摩西管柄囊霉Funneliformis mosseae对正常供水与干旱处理的盆栽枳Poncirus trifoliata实生苗生长、活性氧代谢及抗氧化酶基因表达量的影响。结果表明,7周干旱处理显著降低了根系菌根侵染率。接种摩西管柄囊霉显著促进了干旱处理的枳植株生长,增加了根系体积和叶片相对含水量,显著降低了叶片脯氨酸含量,同时也上调了干旱处理的枳叶片精氨酸脱羧酶基因（PtADC1和PtADC2）和超氧化物歧化酶基因（PtFe-SOD和PtMn-SOD）、过氧化物酶基因（PtPOD）和过氧化氢酶基因（PtCAT1）的表达,因而维持了一个相对更低的活性氧水平（如过氧化氢）,有利于增强植株的抗旱性。