吲哚诱导对茶树抗茶小绿叶蝉的影响

为明确吲哚处理对茶树抗茶小绿叶蝉Empoasca onukii的影响,以龙井43为材料,使用吲哚诱导茶树作为处理,以未处理茶树作为对照,于室内测定茶小绿叶蝉对不同茶树的选择趋向,利用昆虫刺探电位图谱分析茶小绿叶蝉的取食行为,通过气相色谱-质谱联用仪分析吲哚处理对茶树挥发物的影响,并利用实时荧光定量PCR技术测定茶树防御相关基因的表达量。结果表明,在所有时间段茶小绿叶蝉成虫更趋向于取食对照组茶树,至48 h时对照组茶树上的成虫数量是处理组的2.45倍;茶小绿叶蝉3龄幼虫在吲哚处理组茶树上的非取食波（NP波）持续时间占总取食时间的72.15%,在对照组茶树上的非取食波（NP波）持续时间占比则为33.07%,且在对照组茶树上的主要取食波E2波的持续时间是处理组茶树上的3.58倍。相较于对照组茶树,吲哚处理组茶树释放出的挥发物总量显著增加,其中α-法尼烯可以趋避害虫并对天敌有吸引力,且吲哚处理组茶树的防御基因表达量显著升高。表明吲哚处理茶树后,茶小绿叶蝉对其的取食量下降,茶树的挥发物释放量增加且抗性防御基因表达量提高,增强了茶树对茶小绿叶蝉的抗性。     

玫瑰黄链霉菌活性代谢产物RM诱导黄瓜抗病性的信号转导途径

为明确玫瑰黄链霉菌Streptomyces roseoflavus菌株men-myco-93-63活性代谢产物roflamycoin和men-myco-A(简称RM)诱导黄瓜产生抗病性的信号转导途径,采用高效液相色谱法、分光光度法和实时荧光定量PCR法,测定喷施玫瑰黄链霉菌代谢产物RM后黄瓜叶片中水杨酸含量和脂氧合酶活性,以及水杨酸信号转导途径和乙烯茉莉酸信号转导途径的标志基因——NPR1、PR-1a、CTR1的表达量。结果表明,喷施玫瑰黄链霉菌活性代谢产物RM后120 h,黄瓜叶片内水杨酸含量达到最高值,为7.22μg/g,是对照的1.75倍;喷施玫瑰黄链霉菌活性代谢产物RM后12 h,黄瓜叶片内脂氧合酶活性达到最高值,为52.69 U/g,是对照的1.32倍,并且在120 h内整体活性均高于对照;喷施玫瑰黄链霉菌代谢产物RM后,NPR1和PR-1a基因的相对表达量上调,其最大值分别为0.99和1.35,分别是对照的2.24倍和12.09倍,但抑制CTR1基因的表达。推测玫瑰黄链霉菌代谢产物RM通过水杨酸通路、乙烯通路和茉莉酸通路共同诱导黄瓜抗病性信号转导途径。     

心叶烟NPR1基因全长cDNA序列的克隆及表达分析

 NPR1(non-expressor of pathogenesis-related gene 1)基因在拟南芥系统获得抗性中起着关键作用,可调控拟南芥植株广谱抗性的发生。本文报道了从心叶烟中克隆NPR1同源基因(NgNPR1)及其表达特性的研究结果。NgNPR1 cDNA全长2253 bp,编码588个氨基酸。将NgNPR1基因组全长与cDNA序列进行比对发现,NgNPR1基因组DNA含有4个外显子和3个内含子。Southern杂交分析表明,在心叶烟基因组中NgNPR1为单拷贝基因。采用绿色荧光蛋白在洋葱表皮瞬时表达的试验,证明了NgNPR1蛋白在水杨酸诱导时会从细胞质转运到细胞核中。Northern杂交分析发现,NgNPR1基因可以被与植物抗病相关的信号分子如水杨酸、茉莉酸甲酯、过氧化氢和乙烯所诱导。进一步研究发现,植物病原物如赤星病菌、青枯病菌和烟草花叶病毒对心叶烟植株的侵染也会使NgNPR1表达量增加。这些结果表明,NgNPR1基因在心叶烟植株抵御病原物侵染过程中可能起着重要作用。     

茶园间作功能植物对茶小绿叶蝉的调控作用

为研究间作功能植物对茶园主要害虫茶小绿叶蝉的影响,于2017—2018年在福建省泉州市安溪县茶园间作白三叶Trifolium repens L.、金花菜Medicago hispida Gaertn.、金盏菊Calendula officinalis L.,以自然留养杂草茶园为对照,用网捕法采集茶小绿叶蝉并于室内计数,镜检剥查法调查其茶梢着卵量。结果表明,在4种不同生境管理茶园网捕茶小绿叶蝉总个体数及每月个体数均无显著差异;茶小绿叶蝉个体数分别在2017年8月与2018年6月达到高峰。茶梢各节间着卵量不同,主要产卵在顶芽之下第2～5节间,以第4节着卵量最多。间作金盏菊茶园茶小绿叶蝉在茶梢第1节着卵量显著高于间作白三叶和金花菜茶园。不同生境管理茶园茶小绿叶蝉在茶梢第4节着卵量从大到小依次为间作金盏菊(8.13)、间作金花菜(7.50)、自然留养杂草(7.17)和间作白三叶茶园(6.57)。不同时间段4种不同生境管理茶园茶小绿叶蝉在茶梢各节之间的着卵量存在差异,且均在9月上中旬着卵量达到高峰。在茶园间作功能植物,短期内未见其显著降低茶小绿叶蝉网捕量和茶梢着卵量的效应。     

瑞香素对烟草青枯病的防治效果及其作用机理

为探究植物次生代谢产物瑞香素对烟草Nicotiana tabacum青枯病的防治效果及其诱导抗性作用,通过室内和田间试验评价瑞香素对烟草青枯病的防治效果,利用高效液相色谱（high performance liquid chromatography,HPLC）和实时荧光定量PCR（real-time quantitative PCR,RT-qPCR）技术分析其对烟草体内酶活性、木质素含量、激素水平和抗性相关基因表达量的影响。结果表明,叶面喷施瑞香素对烟草青枯病有较好的防治效果,其中2.85 mg/L处理的防治效果最佳,其次是5.70 mg/L和11.40 mg/L处理,接种后14 d的相对防效依次为66.93%、42.52%和48.03%。瑞香素处理可显著提高烟草体内过氧化物酶、苯丙氨酸解氨酶和多酚氧化酶的活性,处理后12 h分别较对照显著提高了3.56倍、1.68倍和1.82倍;瑞香素处理后4 d和7 d烟草根部木质素含量分别较对照显著提高了1.33倍和1.54倍;瑞香素处理后6 h烟草体内茉莉酸和脱落酸含量分别较对照显著提高了35.98%和34.55%,而水杨酸含量较对照显著降低了25.56%。同时,叶面喷施2.85 mg/L瑞香素后显著抑制了茉莉酸途径拮抗基因JAZ3的表达,处理后6 h的表达量较对照下调了9.19倍,同时显著促进病程相关基因PR1的表达。提前喷施2.85 mg/L瑞香素能有效控制田间烟草青枯病发生,相对防效达到52.22%~75.54%,显著高于对照药剂苯并噻二唑的相对防效33.50%~53.00%。表明瑞香素能提升烟草体内酶活性和根部木质素含量,促进抗性基因表达,对田间烟草青枯病有稳定的防治效果,具有开发为防治烟草青枯病的植物激活剂的潜力。     

基于16S rDNA序列的小贯小绿叶蝉共生细菌多样性研究

利用Illumina HiSeq技术,对赤壁、大悟、武汉、咸安和英山5个茶园小贯小绿叶蝉地理种群的成虫共生细菌16SrDNA-V4变异区进行测序,应用Uparse和RDP Classifier等软件统计和分析样本的物种组成、丰度和多样性。5个地区小贯小绿叶蝉成虫的16SrDNA基因序列文库共获得239 645条有效tags,在97%相似阈值下将其聚类为3 403个OTUs。共注释到41个门,116个纲,197个目,272个科,372个属,105个种。5个样本的共生菌在不同分类水平上的组成有所不同,其中在门水平上,主要优势菌为变形菌门Proteobacteria(相对丰度60.6%~97.1%);在纲水平上,相对丰度排名前5位中共有的优势菌为γ-变形菌纲Gammaproteobacteria和α-变形菌纲Alphaproteobacteria;在属水平上,排名前10位中5个样本共有的优势菌为盐单胞菌属Halomonas、希瓦氏菌属Shewanella和沃尔巴克氏体属Wolbachia。小贯小绿叶蝉成虫细菌的Chao指数、Ace指数、Shannon指数和Simpson指数分别为1 065.55~2 841.89,1 130.76~2 914.90,1.07~8.63和0.18~0.99。茶园小贯小绿叶蝉成虫共生细菌多样性比较丰富,不同地理种群的小贯小绿叶蝉细菌群落结构和多样性有差异。本研究结果为进一步研究细菌对小贯小绿叶蝉种群生物学的影响奠定了基础。     

荔枝采后炭疽病的发生情况及对贮藏效果的影响

为了明确广东、海南荔枝采后主要病害及其发生和危害情况，从两省荔枝主产区取果，在常温贮藏条件下观察荔枝采后病害的发生情况，测定荔枝果皮上炭疽病菌潜伏侵染率及其对荔枝采后生理变化和贮藏效果的影响。结果表明：荔枝采收后在28℃下贮藏，炭疽病、霜疫病和酸腐病的发病率分别为95．3％、6．5％和8．2％。炭疽病在采后3—4天开始表现症状，7天后发病率为60．8％～100％，9天后发病率为83．1％～100％，该病害是引起荔枝采后腐烂的主要病害，其病原菌主要来自采前潜伏侵染。从幼果果皮上可分离出潜伏侵染的炭疽病菌，随着果实的生长，果皮上的潜伏侵染率不断上升，到采收时炭疽病菌潜伏侵染率达到90％左右。炭疽病菌潜伏侵染率较高的荔枝果实。其采后呼吸速率、乙烯释放量、果皮丙二醛含量均显著升高，褐变腐烂较快，贮藏效果较差，即使在采后用杀菌剂处理，防治效果也不明显。采收时荔枝果皮上炭疽病菌潜伏侵染率越低，贮藏效果越好。在荔枝果实生长期间喷雾2—3次杀菌剂可显著地降低炭疽病菌潜伏侵染率，减轻荔枝采收后炭疽病的发生严重程度，提高荔枝贮藏效果。     

唑虫酰胺30%悬浮剂防治茶树茶小绿叶蝉药效试验

作者进行了唑虫酰胺30%悬浮剂防治茶树小绿叶蝉药效试验,药后1d,唑虫酰胺30%悬浮剂各处理对茶小绿叶蝉防效在88.9%~91.8%;药后14d,唑虫酰胺30%悬浮剂各处理对茶小绿叶蝉防效在92.8%~93.8%。     

植保无人飞机低容量喷雾在山东设施茶园防治半翅目害虫的应用研究

利用四旋翼植保无人飞机在山东设施茶园防治主要害虫, 研究两种类型的植保无人飞机和传统背负式电动喷雾器施药在不同冠层茶树叶片上雾滴密度?沉积量和覆盖率分布情况以及对茶树主要半翅目害虫的防治效果?结果表明, Ⅰ型植保无人飞机?Ⅱ型植保无人飞机和背负式喷雾器在茶树叶片上平均雾滴密度分别为102.72?50.58个/cm2和29.83个/cm2, 沉积总量分别为0.175?0.371 μL/cm2和53.562 μL/cm2, 雾滴覆盖率均值分别为1.95%?2.52%和19.42%?不同施药器械对茶树各层的雾滴密度?沉积量和覆盖率有显著影响?两种类型植保无人飞机施药雾滴密度总体为茶树树冠上层>中层>底层, 正面>背面?背负式电动喷雾器施药的雾滴沉积量和覆盖率显著高于植保无人飞机, 叶片正面沉积量和覆盖率明显高于叶背面?Ⅰ型植保无人飞机喷施22%氟啶虫胺腈悬浮剂240 mg/L 3 d和5 d后对黑刺粉虱的防治效果为82.20%和71.73%, 该处理对防治茶树黑刺粉虱具有速效性且持效性较好?Ⅰ型植保无人飞机喷施22%氟啶虫胺腈悬浮剂240 mg/L 3 d和5 d后对小贯小绿叶蝉的防效为84.44%和77.25%; Ⅱ型植保无人飞机喷施25 g/L溴氰菊酯乳油60 mg/L 3 d后对小贯小绿叶蝉的防效为98.18%, 具有较好的速效性?植保无人飞机施药对黑刺粉虱的防效优于背负式喷雾器, 但对叶蝉的防效与背负式喷雾器相当?不同施药器械不同药剂处理中瓢虫的数量没有差异?     

Effects of pre‐ and post‐treatment with ethephon on gum formation of peach gummosis caused by Lasiodiplodia theobromae

Peach gummosis, caused by Botryosphaeria spp. fungi, is the process of gum accumulation and exudation in plants. Ethephon (2‐chloroethylphosphonic acid) has profound effects on plants, including enhanced production of secondary metabolites and regulation of plant diseases. This study investigates the effects of application of ethephon before and after inoculation with Lasiodiplodia theobromae on gum formation. Gum formation was promoted by ethephon treatment prior to pathogen inoculation, but inhibited by ethephon applied after the pathogen. The inhibitory effect was counteracted by 1‐methylcyclopropane, which is an ethylene signal inhibitor. 1‐methylcyclopropane also promoted gum formation. Exposure of three isolates of Botryosphaeria to ethephon inhibited mycelial growth. Both treatment methods increased the sugar content at 12 and 24 h post‐inoculation (hpi). However, the sucrose, glucose and fructose contents were significantly higher in shoots with ethephon post‐treatment (application of ethephon after the pathogen inoculation) than those in shoots with ethephon pre‐treatment (application of ethephon prior to pathogen inoculation) at 48 and 72 hpi. The expression of two putative senescence‐related genes, SEN2 and SEN4, were significantly enhanced in pre‐ and post‐treated shoots with ethephon at 24, 48 and 72 hpi. Ethephon application also up‐regulated expression of the pathogenesis‐related protein PR4 while down‐regulating PR1a and PR10. The results show that ethephon has a dual function in regulating gum formation by affecting both the peach shoots and the pathogen.     

The oligopeptide elicitor Pep-13 induces salicylic acid-dependent and -independent defense reactions in potato

The Phytophthora-derived oligopeptide elicitor, Pep-13, originally identified as an inducer of plant defense in the nonhost–pathogen interaction of parsley and Phytophthora sojae, triggers defense responses in potato. In cultured potato cells, Pep-13 treatment results in an oxidative burst and activation of defense genes. Infiltration of Pep-13 into leaves of potato plants induces the accumulation of hydrogen peroxide, defense gene expression and the accumulation of jasmonic and salicylic acids. Derivatives of Pep-13 show similar elicitor activity in parsley and potato, suggesting a receptor-mediated induction of defense response in potato similar to that observed in parsley. However, unlike in parsley, infiltration of Pep-13 into leaves leads to the development of hypersensitive response-like cell death in potato. Interestingly, Pep-13-induced necrosis formation, hydrogen peroxide formation and accumulation of jasmonic acid, but not activation of a subset of defense genes, is dependent on salicylic acid, as shown by infiltration of Pep-13 into leaves of potato plants unable to accumulate salicylic acid. Thus, in a host plant of Phytophthora infestans, Pep-13 is able to elicit salicylic acid-dependent and -independent defense responses.     

Expression of δ-endotoxin Cry1EC from an inducible promoter confers insect protection in peanut (Arachis hypogaea L.) plants

BACKGROUND: Spodoptera litura (F.) is a polyphagous foliage insect and a major pest on peanut (Arachis hypogaea L.). Constitutive expression of δ‐endotoxin Cry1EC gives protection against S. litura, as reported earlier. In this study, insect bites and salicylic acid induced high‐level expression of Cry1EC was achieved in peanut. In order to achieve this, the expression of pathogenesis responsive promoter PR‐1a was enhanced by placing it downstream of the CaMV35S promoter in the pCAMBIA 1300 backbone. The resultant promoter CaMV35S(r)PR‐1a expressed a high level of insecticidal δ‐endotoxin Cry1EC. The Gus expression under the control of CaMV35S(r)PR‐1a served as a convenient marker for evaluation of promoter response to different treatments. RESULTS: Transgenic events that showed a very low level of uninduced expression and no expression in seeds were selected. The Cry1EC expression in leaves increased nearly eightfold in the selected event, following induction by salicylic acid. Both the salicylic‐acid‐treated and the S. litura‐bitten leaves showed the highest expression after 2 days. Leaves from salicylic‐acid‐induced transgenic plants caused 100% mortality of S. litura at all stages of larval development. CONCLUSION: The results suggest that high expression of inducible promoters provides a good strategy for the development of safer transgenic food and feed crops. Copyright © 2010 Society of Chemical Industry