赤星病胁迫对不同抗性烟草品种光合作用和叶绿素荧光特性的影响

以不同烟草赤星病抗性品种JYH(抗病品种)和CBH(感病品种)为材料,在盆栽试验条件下,调查不同烟草赤星病胁迫程度(轻度胁迫、中度胁迫和重度胁迫)对光合色素含量、光合作用参数和叶绿素荧光动力学特征的影响。结果显示:1)烟草赤星病胁迫导致2个品种的叶绿素a、叶绿素b、总叶绿素和类胡萝卜素含量均呈下降趋势,且JYH的降幅小于CBH。2)除JYH的净光合速率在轻度胁迫下有所增加外,JYH、CBH的净光合速率、气孔导度因烟草赤星病胁迫而降低。2个品种的胞间CO2浓度、气孔限制值变化具有明显差异,烟草赤星病胁迫导致CBH的胞间CO2浓度上升,气孔限制值则明显下降,这与JYH在重度胁迫下的变化趋势一致。而JYH的胞间CO2浓度在轻度、中度胁迫降低,气孔限制值则呈上升趋势。3)烟草赤星病胁迫下,2个品种的初始荧光(F0)、非光化学猝灭系数(NPQ)均有所增加。重度胁迫下,JYH、CBH的F0分别比对照增加16.5%、34.48%,NPQ分别上升95.54%、137.45%,差异均达到显著水平。而各品种的最大荧光(Fm)、可变荧光(Fv)、PSⅡ最大光化学效率(Fv/Fm)、PSⅡ潜在活性(Fv/F0)、光化学淬灭系数(qp)、PSⅡ实际光化学效率(ΦPSⅡ)在烟草赤星病胁迫下均呈下降趋势,降幅表现为JYHCBH。研究结果表明,JYH的光合色素、光合作用和叶绿素荧光特性受烟草赤星病胁迫影响小于CBH,维持较高的光合性能是对烟草赤星病具有较强抗性的生理基础。     

干旱胁迫对转BnDREB1-5基因烟草光合特性的影响

检测了干旱胁迫对烟草(Nicotiana tabacum)叶片光合特性、叶绿素、类胡萝卜素和可溶性糖含量的影响.结果显示,干旱胁迫后转基因烟草的初始荧光(Fo)、非光化学猝灭系数(NPQ)增加幅度明显低于野生型烟草;其它荧光参数降低幅度明显小于野生型烟草;虽然干旱胁迫后两个烟草样品的光合参数、叶绿素含量都有所降低,但转基因烟草的降低幅度明显小于野生型烟草;干旱胁迫后,转基因烟草可溶性糖含量明显高于野生型烟草.表明BnDREB1-5转录因子具有抗干旱胁迫的作用.     

转Bt基因棉花杀虫晶体蛋白的表达及光合特性的研究

转Bt基因棉花(GK、ZK)及非Bt基因棉花(CZ)杀虫晶体蛋白表达及光合特性的研究表明,杀虫晶体蛋白在转Bt基因棉花GK与ZK中的表达总量及在各器官中的分配均有所不同．转Bt基因棉花叶片的净光合速率的光响应与常规棉有所不同．转Bt基因棉花GK与ZK叶片的叶绿素含量、净光合速率、蒸腾速率的日变化有明显的不同,而胞间二氧化碳浓度、气孔限制值、叶温的日变化趋势则基本一致．胞间二氧化碳浓度的日平均值在两转Bt基因棉花间的差异达显著水平,而其它各指标在不同处理间的差异均未达显著．     

草莓品种灰霉病抗性田间鉴定

调查了160个草莓品种对灰霉病的田间抗、感病情况,结果表明,草莓品种对灰霉病均无免疫能力,但品种之间的抗、感病程度有差异,抗病性较强的品种为硕蜜,感病率为4.8%,感病率最高的品种为绥陵,达66.7%。筛选出硕蜜、83-11、益芳、明宝、83-21等7个品种(系)表现为高抗(HR)。高抗品种中,生长势以中偏强者、株态以半直立至半开张者较多。     

镉胁迫对烟草叶激素水平、光合特性、荧光特性的影响

采用不同浓度镉胁迫（0mg/L、5mg/L、25mg/L、50mg/L）的水培试验,分别用感应耦合等离子体、紫外分光光度计、酶联免疫法、光合作用仪、叶绿素荧光分析仪研究了镉胁迫3d对烟草(Nicotiana tabacum L.)叶内镉含量、IAAO活性、激素（IAA、ABA、GA3、Z）水平、光合特性及荧光特性的影响。结果显示烟草叶内镉含量随着胁迫浓度增加而显著增加;IAAO活性在镉为5mg/L时下降,然后随着镉浓度增加而显著上升;IAA含量在镉为5mg/L时上升,但随着镉浓度增加而显著下降;ABA含量随着镉浓度上升而增加;GA3含量在5mg/L时上升,然后随着镉浓度增加而显著下降;Z含量随着镉浓度增加而显著下降;叶绿素a、叶绿素b和类胡萝卜素含量均随镉胁迫浓度增大而显著减少;光合参数（Pn、Gs、Ci、Tr）及荧光参数（ Fv/ Fo、Fv/Fm、ФPSⅡ、qP）均随镉胁迫浓度增大而下降。     

烟草NtNHX1 3基因的克隆及表达特性

以栽培烟草‘云烟87’为材料,通过同源克隆方法分离了烟草NHX（Na⁺,K⁺/H⁺ exchanger）基因NtNHX1 3。结果表明：NtNHX1 3基因CDS长度1 617 bp,编码蛋白质长度为538 aa,蛋白理论等电点为8.67,分子量为59.34 kD。预测NtNHX1 3属于膜蛋白,含有11个跨膜区,且含有NHX类蛋白保守位点氨氯吡嗪咪结合位点。进化树分析显示,NtNHX1 3与菊苣、野菊花NHX遗传距离最近。qRT PCR组织特异性表达分析表明,NtNHX1 3在烟草叶片中表达量最高,根、茎、花中也有表达;盐胁迫处理后NtNHX1 3基因的表达上调,表明该基因参与了盐胁迫反应;打顶初期NtNHX1 3基因的表达呈逐渐上调的趋势,与该时期钾含量逐渐升高相吻合。研究推测,NtNHX1 3具有将钾离子从细胞质转运至液泡的功能。     

不同光环境下烟草光合特性及同化产物的积累与分配机制

为了解烟草光合特性与光合作用同化产物的积累与分配对不同光环境的适应,以盆栽烟草为试验对象,于人工气候室中设置3种光照强度[遮阴(400±15)～(500±15) μmol·m^-2·s^-1;自然光强:(800±15)～(1000±15) μmol·m^-2·s^-1;高光强:(1500±15)～(1800±15) μmol·m^-2·s^-1]系统研究光照条件对烟草光合特性及光合作用同化产物在烟株-土壤系统分配的影响.结果表明: 随着光照强度的降低,烟草各组分生物量逐渐减小,根冠比降低.净光合速率(P_n)、气孔导度(g_s)、蒸腾速率(T_r)均随光照强度的减弱呈下降趋势,胞间CO₂浓度(C_i)升高;在强光条件下烟草最大净光合速率(A_max)、光饱和点(LSP)、光补偿点(LCP)、暗呼吸速率(R_d)均达到最大值,弱光条件则具有较大的表观量子效率.光照强度影响烟草对¹³C的吸收、积累与分配,弱光条件下,烟草富集的¹³C进入到根部的比例明显较少,更多的分配到地上部.由此可知,外界光环境的变化不仅显著影响烟草叶片的光合特性与生物量积累,也使光合碳在烟株-土壤系统的分配格局发生变化.     

转金属硫蛋白突变体αα的烟草具有较高的重金属抗性

金属硫蛋白（ＭＴ）有α、β两个结构域,其中α结构域优先结合Ｃｄ＾２＋、Ｈｇ＾２＋以α结构域代替β结构域而得到的αα突变体在大肠杆菌构建及表达后,体外实验证明其具有同天然ＭＴ相似的稳定性,但结合重金属的能力稍强于天然ＭＴ。将αα突变体转入植物表达载体,以ＣａＭＶ３５Ｓ为启动子,利用根癌土壤杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍ ｔｕｍｅｆａｃｉｅｎｓ（Ｓｍｉｔｈ ｅｔ Ｔｏｗｎｓｅｎｄ）Ｃｏｎｎ）介导的叶     

茶树转录因子CsbHLH137基因鉴定及光合特性与生物钟响应分析

基于茶树基因组数据,该研究以茶树‘蒙山9号’cDNA为模板,采用PCR扩增技术,成功克隆得到开放阅读框（ORF）长度为1 023 bp,编码340个氨基酸的茶树bHLH家族转录因子基因,命名为CsbHLH137（登录号为OL332046）。蛋白序列特征分析表明,CsbHLH137转录因子含有HLH结合功能域,且bHLH蛋白功能之一为通过生物钟组成结构域调控对光的反应和相互作用。该蛋白有4个无序化区域,包含有20个磷酸化位点,相对分子量为38.59 kD,理论等电点为5.59,属于亲水性蛋白。CsbHLH137转录因子蛋白二级结构以α-螺旋和随机卷曲为主。对不同时间点的茶树叶片进行气孔开度分析和光合参数测定结果显示,与黑暗处理相比,光照处理在调节茶树叶片气孔宽度方面更为明显,光合参数G_s、C_i和T_r在白天波动较大,夜间维持较稳定状态。实时荧光定量PCR技术检测表明,茶树CsbHLH137转录因子基因在白天的表达量高且出现峰值,在夜间维持平稳的低表达水平。研究推测,茶树CsbHLH137转录因子基因为DELLA蛋白的应答基因...     

转玉米pepc基因水稻的某些光合特性

测定转玉米pepc基因水稻和未转化粳稻品种Kitaake在分蘖初期、分蘖盛期、拔节期、始穗期、齐穗期、成熟期和剑叶不同生长时期的光合特性动态变化的结果表明:转玉米pepc基因水稻的磷酸烯醇式丙酮酸羧化酶(PEPCase)活性和净光合速率(Pn)在不同时期各不相同,相对于Kitaake而言,均有不同程度的提高;剑叶完全展开时的PEPCase活性和Pn提高最明显.     

外源钙和茉莉酸甲酯诱导番茄植株抗灰霉病研究

以‘辽园多丽’番茄幼苗为材料,研究了经钙(Ca)、钙螯合剂(EGTA)和茉莉酸甲酯(MeJA)处理后接种番茄灰霉病幼苗叶片的病情指数、活性氧(H2O2、O2.-)含量和过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、过氧化物酶(POD)活性的变化。结果显示:(1)Ca、MeJA、MeJA+Ca处理番茄幼苗的灰霉病发病率分别比对照显著降低32.5%、38.0%和54.5%,而MeJA+Ca处理又显著低于Ca、MeJA处理32.6%和15.3%;MeJA+EGTA处理高于MeJA处理30.3%,但低于EGTA处理13.1%;Ca处理低于EGTA处理34.2%。(2)Ca、MeJA及MeJA+Ca处理番茄幼苗叶片中活性氧积累量高于对照,MeJA+Ca处理又高于Ca、MeJA处理;但MeJA+EGTA处理活性氧积累量低于MeJA处理,而高于EGTA处理;Ca处理的活性氧含量高于EGTA处理。(3)Ca、MeJA及MeJA+Ca处理幼苗叶片的SOD、CAT、POD的活性均比对照提高,且以MeJA+Ca处理最高;而MeJA+EGTA处理抗氧化酶活性低于MeJA处理,但高于EGTA处理;Ca处理抗氧化酶活性高于EGTA处理。研究表明,钙在茉莉酸甲酯诱导番茄抗灰霉病过程中具有重要调节作用,这种作用与钙促进茉莉酸甲酯诱导番茄活性氧积累和抗氧化酶活性有关。     

灰葡萄孢菌过氧化物酶体及细胞核的荧光蛋白标记

[目的]对灰葡萄孢菌(Botrytis cinerea)的细胞核和过氧化物酶体进行荧光蛋白标记,为研究其生长发育和侵染过程中细胞结构和细胞器动态提供基础.[方法]以绿色荧光蛋白(GFP)和红色荧光蛋白(DsRED、mCherry)为报告基因,利用根癌农杆菌介导转化(Agrobacterium tumefaciens m...     

嗜线虫致病杆菌抑制灰葡萄孢的效应

[背景] 灰葡萄孢（Botrytis cinerea）是引起葡萄采后病害的主要病原菌之一,严重影响葡萄的贮期和品质,给葡萄产业带来极大损失。利用拮抗微生物抑制采后病原菌生长已逐渐成为防治葡萄采后灰霉病的重要手段。[目的] 利用昆虫病原线虫共生细菌广谱高效的抑菌特性,从现有共生细菌资源中筛选对灰葡萄孢具有高拮抗作用的菌株,为葡萄采后灰霉病的抑制提供新的材料和研究方向。[方法] 通过平板对峙培养法和菌丝生长速率法分离筛选拮抗共生细菌,并对优选的高效拮抗共生细菌进行16S rRNA基因序列进化分析,采用扫描电镜观察其对灰葡萄孢菌丝生长的影响,利用损伤接种法对红地球葡萄防治效果进行验证。[结果] 初步分离筛选共获得9株拮抗菌,复筛与复测得到一株抑菌效果显著的共生细菌（命名为ALL）,经进化分析其为嗜线虫致病杆菌（Xenorhabdus nematophila）,其16S rRNA基因序列的Genbank登录号为MW488402,与菌株Xenorhabdus nematophi la NC116聚于同一分支,相似性达99.79%。扫描电镜观察该菌株导致灰葡萄孢菌丝扭曲变形、表面皱缩、失水塌陷,该菌株发酵（36 h）上清液浓度为1%时对灰葡萄孢菌丝抑制率达44.5%。在葡萄常温防效实验中,与对照组比较,ALL菌株发酵上清液对灰霉菌防治效果较好,3 d后防效为63.50%。[结论] 本研究应用昆虫病原线虫共生细菌生物防治葡萄贮期灰霉病,筛选出一株高效拮抗灰葡萄孢的昆虫病原线虫共生细菌,而且其上清液对灰葡萄孢具有良好的抑制效果,为生物防治贮期葡萄灰霉病提供了新的生物材料和相关研究基础。     

Induced proteome of Trichoderma harzianum by Botrytis cinerea

As a notable biocontrol agent, Trichoderma harzianum can antagonize a diverse array of phytopathogenic fungi, including Botrytis cinerea, Rhizoctonia solani and Fusarium oxysporum. Elucidating the biocontrol mechanism of T. harzianum in response to the pathogens enables it to be exploited in the control of plant diseases. Two-dimensional gel electrophoresis (2-DE) was performed to obtain secreted protein patterns of T. harzianum ETS 323, grown in media that contained glucose, a mixture of glucose and deactivated B. cinerea mycelia, deactivated B. cinerea mycelia or deactivated T. harzianum mycelia. Selected protein spots were identified using liquid chromatography–tandem mass spectrometry (LC–MS/MS). Ninety one out of 100 excised protein spots were analyzed and some proteins were sequence identified. Of these, one l-amino acid oxidase (LAAO) and two endochitinases were uniquely induced in the media that contained deactivated B. cinerea mycelia as the sole carbon source. Activities of the cell wall-degrading enzymes (CWDEs), including β-1,3-glucanases, β-1,6-glucanases, chitinases, proteases and xylanases, were significantly higher in media with deactivated B. cinerea mycelia than in other media. This finding suggests that the cell wall of B. cinerea is indeed the primary target of T. harzianum ETS 323 in the biocontrol mechanism. The possible roles of LAAO and xylanase were also discussed.     

利迪链霉菌K2对灰霉病菌的抑菌效果及抑菌物质鉴定

灰霉病是多种经济作物生产过程以及果蔬储藏运输中常见的病害,链霉菌能产生丰富的次级代谢产物,对灰霉病菌具有较好的抑制效果。【目的】筛选出更高效、功能更多的链霉菌,为针对灰霉病的生防菌剂的研发提供优良菌种。【方法】采用管碟法对菌株K2进行液体培养基的筛选及培养液活性的测定;双皿对峙等2种方法进行产挥发性物质对灰霉病菌的抑菌活性测定;通过16S rRNA基因序列分析进行菌株K2鉴定;高效液相色谱及液相色谱-质谱2种方法对培养液活性成分进行定性验证;顶空固相微萃取-气质联用对菌株K2产生的挥发性物质成分进行检测及鉴定。【结果】菌株K2在液体培养基A中产生的次级代谢物对苹果腐烂病菌、苹果轮纹病菌、核盘菌、杨树溃疡病菌和烟草赤星病菌等多种植物病原真菌均具有较强的抑制作用;K2产生的挥发性物质对灰霉病菌的抑制率达100%,且抑制效果与挥发性物质的量有关;菌株K2与利迪链霉菌(Streptomyces lydicus)亲缘关系非常接近,相似性为99%;培养液活性成分中含有谷氏菌素、丰加霉素和纳他霉素;在挥发性物质成分中发现了烯类、醇类、酯类及烷烃类等30种挥发性物质,其中含量较多的物质分别是2-met...     

Purification and characterization of two isozymes of polygalacturonase from Botrytis cinerea. Effect of calcium ions on polygalacturonase activity

The phytopathogenic fungus Botrytis cinerea produces a set of polygalacturonases (PGs) which are involved in the enzymatic degradation of pectin during plant tissue infection. Two polygalacturonases secreted by B. cinerea in seven-day-old liquid culture were purified to apparent homogeneity by chromatography. PG I was an exopolygalacturonase of molecular weight 65 kDa and pI 8.0 and PG II was an endopolygalacturonase of 52 kDa and pI 7.8. Enzymatic activity of PG I and PG II was partially inhibited by 1 mM CaCl₂, probably by calcium chelation of polygalacturonic acid, the substrate of the enzyme.     

Evaluation of formulations of Bacillus licheniformis for the biological control of tomato gray mold caused by Botrytis cinerea

Bacillus licheniformis N1, which has previously exhibited potential as a biological control agent, was investigated to develop a biofungicide to control the gray mold of tomato caused by Botrytis cinerea. Various formulations of B. licheniformis N1 were developed using fermentation cultures of the bacteria in Biji medium, and their ability to control gray mold on tomato plants was evaluated. The results of pot experiments led to the selection of the wettable powder formulation N1E, based on corn starch and olive oil, for evaluation of the disease control activity of this bacterium after both artificial infection of the pathogen and natural disease occurrence under production conditions. In plastic-house artificial infection experiments, a 100-fold diluted N1E treatment was found to be the optimum biofungicide spray formulation. This treatment resulted in the significant reduction of symptom development when N1E was applied before Bo. cinerea infection, but not after the infection. Both artificial infection experiments in a plastic house and natural infection experiments under production conditions revealed that the N1E significantly reduced disease severity on tomato plants and flowers. The disease control value of N1E on tomato plants was 90.5% under production conditions, as compared to the 77% conferred by a chemical fungicide, the mixture of carbendazim and diethofencarb (1:1). The prevention of flower infection by N1E resulted in increased numbers of tomato fruits on each plant. N1E treatment also had growth promotion activity, which showed the increased number of tomato fruits compared to fungicide treatment and non-treated control and the increased fruit size compared the non-treated control under production conditions. This study suggests that the corn starch-based formulation of B. licheniformis developed using liquid fermentation will be an effective tool in the biological control of tomato gray mold.     

甲基营养型芽孢杆菌拮抗玉蜀黍尾孢菌、链格菌和灰葡萄孢菌及环脂肽分析

[目的]探究甲基营养型芽孢杆菌(Bacillus methylotrophicus)对植物病原菌玉蜀黍尾孢菌(Cercospora zeae-maydis Tehon et Daniels)、链格菌(Alternaria alternate)和灰葡萄孢菌(Botrytis cinerea)的拮抗作用并鉴定抗菌物质,为其...     

Suppression of Botrytis cinerea sporulation by Clonostachys rosea on rose debris: a valuable component in Botrytis blight management in commercial greenhouses

Botrytis blight, caused by Botrytis cinerea (Bc), is an important disease on roses grown in plastic greenhouses in Brazil. Biocontrol with Clonostachys rosea (Cr) applied to leaves and crop debris to reduce pathogen sporulation can complement other control measures for disease management. Two experiments, each with a rose cultivar, were conducted in a plastic greenhouse. For ‘Red Success,’ four treatments were compared: (1) control; (2) fortnightly sprays of Cr; (3) weekly sprays of mancozeb; and (4) weekly sprays of either Cr or mancozeb to the lower third of the plants and the debris. For ‘Sonia,’ treatment 4 was not included. Samples were taken from debris (leaves and petals) at ten 15-day intervals and plated on PCA medium. Sporulation of fungi and incidence of Botrytis blight on buds were assessed. For both cultivars, C treatments significantly (P=0.05) reduced Bc sporulation. However, disease incidence was not consistently reduced, probably because the applications of C. rosea started when Botrytis blight epidemic was advanced and no sanitation practices were performed on nontreated plots. From the present and previous studies, continuous application of Cr on debris, associated with sanitation practices, has the potential to reduce Bc sporulation and disease incidence in the buds.     

蜡质芽胞杆菌XF的发酵条件优化及对番茄灰霉病的防治效果

由灰葡萄孢菌(Botrytis cinerea)引起的番茄灰霉病是一种土传病害,近年来施用生防菌被认为是一种有效的防治手段.以灰葡萄孢菌为指示菌,生物量和抑菌率为指标,采用单因素试验、正交优化试验相结合的方法,优化蜡质芽胞杆菌XF株的发酵培养基和发酵条件,从而提高菌株XF对番茄灰霉病菌的抑制效果,并于室内盆栽条件下探讨...