玉米抵御玉蜀黍尾孢菌侵入的生理机制

为深入探讨玉米抗灰斑病的机制,以抗/感玉米灰斑病自交系78599-1、OH43Ht N和掖478、K12为材料,采用比色皿法和RT-PCR法相结合研究了接种玉蜀黍尾孢菌毒素后玉米叶片中防御酶活性、丙二醛(MDA)含量及防御酶合成相关基因SOD、CAT、APX和GR的表达量。结果显示,接种该病菌毒素后,抗、感材料叶片中防御酶活性均升高,多数在接种7 d时达到峰值,且抗病材料防御酶活性峰值高于感病材料;供试材料叶片中MDA含量均降低,且抗病材料低于感病材料;供试材料叶片中SOD、CAT和GR的表达量均上升,其中CAT的表达量在78599-1、掖478和K12接种5d时达到峰值,灰度值分别为228.67、161.33和178.00,与其酶活性变化趋势一致;SOD和GR的变化规律与其酶活性变化不一致;APX的表达量仅在OH43Ht N中上升,接种7 d后达到峰值。表明抗病材料调控防御酶活性的能力强,防御酶基因的表达与其酶活性变化存在关联性。     

串珠镰刀菌引起玉米穗粒腐病防御酶变化及其电镜观察

 对串珠镰刀菌(Fusarium moniliforme)侵染引起玉米穗粒腐病的防御酶活性变化和病原菌侵染过程进行研究。采用人工接菌的方法,分别对抗(Bt鄄1)、感(掖478)玉米材料进行接种,取抗、感材料间隔24 h 的6 个时间段接菌部位的苞叶组织,分析玉米植株感病后部分防御酶、同工酶谱的动态变化,并用扫描电镜对病原菌入侵植株过程进行组织病理学观察。扫描电镜观察发现,菌丝首先要经过1 ~ 3 d 生长后,大约在72 h 左右开始侵入气孔,并且随着时间的推移,侵入气孔的菌丝量逐渐增多。这说明病原菌是直接通过气孔侵入寄主苞叶组织。同时,玉米受串珠镰刀菌侵染后,苯丙氨酸解氨酶 (PAL)和过氧化物酶(POD)的活性都是先上升后下降,在感病材料Ye478 中PAL 的活性要比抗病材料Bt鄄1 中增加的更快、更高;同样对于POD 来说,在感病材料Ye478 中的活性要比抗病材料Bt鄄1 中的高,但变化趋势在2 个材料中相似;而丙二醛(MDA)的含量则相反,在感病材料Ye478 中的活性要比抗病材料Bt鄄1 中的低;对POD 同工酶酶谱分析,2 个材料都增加了3 ~ 4 个条带,没有明显的区别,这说明玉米感病后会通过增加POD 的活性来抵御外源病菌的侵入。总体而言PAL和POD 活性水平与材料抗性呈负相关;MDA 与材料抗性呈正相关关系。对玉米植株感病后防御酶活性变化的分析和病原菌入侵寄主的电镜观察结果,可为深入研究玉米穗粒腐病抗病机制和抗病育种提供参考。     

玉米生产品种和部分自交系对鞘腐病的抗性筛选初报

玉米鞘腐病[Fusarium proliferatum(Mats.)Nirenberg]常在玉米生育中、后期发生,近年有逐年加重的趋势,对玉米产量造成不同程度的损失。为明确目前生产上推广的玉米品种对鞘腐病的抗性能力,预测该病害的发生、发展趋势,分别于2009年和2010年采用人工接种的方法,对华北和东北玉米产区生产上主推的玉米材料进行了鞘腐病抗病资源筛选。筛选结果表明,目前生产上推广的大多数玉米品种对鞘腐病均具有较好的抗性。     

生姜抗姜瘟病品种资源鉴定及其抗病生化机制

为鉴定生姜抗姜瘟病品种并明确其抗病生化机制,采用茎基部注射法和土壤灌根法接种测试22个生姜品种对姜瘟病的抗性水平,以高抗品种CDZIN012和高感品种CDZIN064为材料接种假茄科雷尔氏菌Ralstonia pseudosolanacearum,观察接种后抗、感品种的叶片气孔参数、蜡质含量等组织结构的变化,测定接种后过氧化物酶(peroxidase,POD)、多酚氧化酶(polyphenol oxidase,PPO)、过氧化氢酶(catalase,CAT)、超氧化物歧化酶(superoxide dismutase,SOD)活性,O₂^-、H₂O₂含量及病程相关蛋白几丁质酶和β-1,3-葡聚糖酶活性的变化。结果显示,22个生姜品种抗性差异明显,其中表现为高抗、中抗、轻抗、感病和高感的品种分别为1、4、4、7和6个;茎基部注射法接种后生姜植株发病更稳定,更适合生姜品种抗性鉴定;感病后抗病品种叶片气孔密度为36.8个/mm²,比感病品种少(46.9个/mm²),...     

杨树体内苯丙烷代谢与其对溃疡病抗性的关系

 本文采用组培愈伤组织接种法研究了不同抗病类型的杨树品种受溃疡病菌(Dothiorella gregaria Sacc.)侵染后苯丙氨酸解氨酶活性以及绿原酸和木质素含量的变化与抗病性的关系。纳果发现,受病菌侵染后各品种的苯丙氨酸解氮酶活性都有一个明显消长过程,但抗病品种的酶活性升高的幅度大(为感病品种的2倍),持续的时间也长;伴随酶活性的变化,各品种的绿原酸含量也发生明显变化,与感病品种相比,抗病品种的绿原酸不仅出现得早,而且积累的量也大(为感病品种的三倍);木质素的增量也与品种的抗病性呈正相关。上述结果表明,杨树对溃疡病的抗性与苯丙氨酸解氨酶的活性密切相关,该酶在抗病中的作用是通过其控制的代谢产物得以表达。     

枯草芽孢杆菌TR21对香蕉抗病相关基因表达的诱导作用

广谱抗真菌枯草芽孢杆菌Bacillus subtilis菌株TR21在温室和大田对香蕉枯萎病具有较好的防效,其机制已证明与诱导香蕉产生系统抗性有关。本文以巴西蕉（Musa AAA Cavendish subgroup cv.Brazil）为材料,利用半定量RT-PCR法,以香蕉25S rRNA基因为内标,研究灌根接种菌株TR21后对香蕉根系4种抗病相关基因表达的影响。结果表明,PAL、POD、PR-3和PR-1基因在接种后表达水平均表现上调趋势,但PAL和POD基因的表达增幅明显高于PR-3和PR-1基因。PAL和POD基因在接种后12 h表达量最高,与TR21在香蕉根部的定殖规律表现出一致性。系统诱导抗性是枯草芽孢杆菌TR21防治香蕉枯萎病的机制之一。     

黑痣病菌毒素对马铃薯幼苗生理生化抗性相关物质的诱导

为了探讨马铃薯抗黑痣病机制,采用毒素接种脱毒组培苗的方法,研究了黑痣病菌毒素对马铃薯不同抗性品种幼苗体内病程相关蛋白、木质素及游离脯氨酸含量的影响及其与抗黑痣病的关系。毒素处理后,感病品种大西洋和抗病品种底西芮幼苗几丁质酶和β-1,3-葡聚糖酶活性均明显升高,36 h升高幅度均达到最大,感病品种升幅大于抗病品种。底西芮和大西洋木质素及游离脯氨酸含量也大幅增加,但抗病品种的增加幅度大于感病品种。研究表明,黑痣病菌毒素诱导几丁质酶和β-1,3-葡聚糖酶活性升高以及木质素和游离脯氨酸含量增加是马铃薯幼苗抵抗毒素胁迫的内在机制;木质素、游离脯氨酸含量与品种抗病性呈正相关,而几丁质酶、β-1,3-葡聚糖酶活性变化则不是引起品种抗性差异的主要原因。     

牛蒡低聚果糖诱导烟草对烟草花叶病毒的抗性

在室内条件下,研究了牛蒡低聚果糖对烟草花叶病毒抗性的影响及其影响机制。结果表明,在牛蒡低聚果糖处理叶（第三叶）中,接种烟草花叶病毒24h时对照处理叶片中该病毒含量大约是牛蒡低聚果糖处理叶片中的2.6倍,相对防效达61%;在未处理叶（第四叶）中,接种烟草花叶病毒24h时对照处理叶片可检测到TMV-CP的表达,而牛蒡低聚果糖处理叶则检测不到TMV-CP的表达。通过对抗病相关基因的检测发现,牛蒡低聚果糖处理和接种烟草花叶病毒都可诱导抗病相关基因（包括PR-1a、PR-2、PR-3、PI-1、PI-2和PAL）在烟草局部叶和系统叶中大量表达。这些结果表明,牛蒡低聚果糖可诱导烟草对烟草花叶病毒产生抗性,其作用机制可能在于诱导了抗病相关基因在植株体内的表达,增强了植株的系统抗性。     

丁香酚对烟草抗烟草花叶病毒的诱导作用初探

为探讨植物源化合物丁香酚对烟草抗烟草花叶病毒(TMV)的诱导作用,初步研究了丁香酚对接种TMV的烟草叶片中叶绿素和丙二醛(MDA)含量,防御酶苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)和超氧化物岐化酶(SOD)活性,以及病程相关蛋白基因 PR-1 、PR-3 、PR-5 表达的影响。结果表明:丁香酚处理可促进叶绿素的合成,减轻烟草体内MDA的积累,提高PAL、POD和SOD的活性,增强 PR-1 、PR-3 、PR-5 基因的表达。表明丁香酚可提高植物的诱导抗病性,进而减轻TMV对烟草的危害。     

大白菜幼苗叶片抗氧化酶系统对黑腐病菌入侵的反应

以大白菜抗病品种秦白2号和感病品种06J31为材料,研究其接种黑腐病菌种YL-17后保护酶活性、超氧阴离子和MDA含量的变化。结果显示:接种后不同抗性的材料总体上表现为超氧阴离子(O2-.)含量先降后升,SOD活性逐步降低,POD活性MDA含量逐步升高,CAT活性则二者变化不一致(抗性材料表现为升-降-升,感性材料表现为先降后升)。由此表明:黑腐病菌侵染大白菜后,抗病材料与感病材料相比,能迸发较多的O2-.并刺激抗氧化物酶SOD和POD活性升高,使MDA含量处于较低的水平,有利于提高秦白2号抗黑腐病和抗氧化能力。     

Effects of a lectin-like protein isolated from Acacia farnesiana seeds on phytopathogenic bacterial strains and root-knot nematode

Acacia farnesiana lectin-like protein (AFAL) showed bacterioestatic effects against Xanthomonas axonopodis pv. passiflorae (Gram-negative) and Clavibacter michiganensis michiganensis (Gram-positive), with the latter being more sensitive. This effect is probably due to the ability of AFAL to interact with the bacterial cell wall where we observed that AFAL induced macroscopic change. The maximum bacterial growth inhibition was approximately 78% when incubated with Gram-negative strains, and as high as 92% percent for the Gram-positive one. The antibacterial effect of flavonoids (rutin, quercetin and morin) was also observed using low concentrations against both bacterial strains. Prior incubation of both with AFAL at high concentrations increases the inhibitory effect of flavonoids on bacterial growth. The potential use of AFAL as a control agent against the root-knot nematode Meloidogyne incognita was investigated as well, showing anti-nematode properties involving both egg hatching and motility. In the juvenile second-stage, AFAL showed reduction in larval mobility when measured against a control group. The results suggest that AFAL is effective against M. incognita and could be used as a component of integrated pest management programs. These data also suggest that lectins probably play a role in plant defense not only against invertebrate phytopathogens, herbivores and fungi but also against bacteria.     

枯草芽胞杆菌对离体荔枝果实霜疫霉病、炭疽病的防治效果

为了进一步明确从土壤中筛选得到的拮抗细菌OR-1、OR-2、OR-3、ON-6的分类地位和生防效果,采用形态学观察、理化特性结合分子生物学方法,鉴定这4株细菌皆为枯草芽胞杆菌Bacillussubitilis.拮抗菌对荔枝霜疫霉菌、炭疽菌菌丝生长均具有明显抑制作用,ON-6菌株的发酵液对菌丝生长的抑制率最高,分别为92.34％和70.36％,其次是OR-1菌株.拮抗菌处理组褐变指数均小于对照组以及杀菌剂处理组,且差异达显著水平(P＜0.05),OR-1菌株防褐变效果最好.常温下,拮抗菌发酵液对离体鲜果病害防治效果优于对照组和杀菌剂处理组,其中ON-6菌株的防治效果最佳;4℃低温储藏处理40天后,拮抗菌发酵液处理组的防治效果高于对照和杀菌剂处理组,且差异显著(P＜0.05),以ON-6和ON-3菌株的防治效果最佳.表明拮抗菌发酵液对荔枝鲜果上的霜疫霉菌和炭疽病菌有一定的抑制作用.     

Agrobacterium-mediated transformation efficiency is altered in a novel rice bacterial blight resistance cultivar and is influenced by environmental temperature

Y73 is a progeny of asymmetric somatic hybridization from an elite japonica rice cultivar (Dalixiang) and a wild rice (Oryza meyeriana), which shows high resistance to bacterial blight. It has a similar genetic background to its recurrent parent, Dalixiang, but Y73 has a high resistance to both bacterial blight and Agrobacterium. The transformation efficiency of Y73 was 35.7%, while Dalixiang had higher transformation efficiency (71.2%) under the same co-cultivation temperature (25 °C). These results indicate that the resistance to Agrobacterium tumefaciens in Y73 was linked with its characteristic of bacterial blight resistance, and was obtained from its donor parent, O. meyeriana. Further studies also showed that the resistance to A. tumefaciens was weakened at a lower temperature (20 °C). To study its molecular mechanism, the expression levels of genes associated with Agrobacterium-mediated transformation (OsMPKs, OsVIP1s and OsPR1s) in rice were investigated by Quantitative real-time PCR (qRT-PCR) analysis. The expression levels of OsMPK genes remained unchanged at different temperatures, while all OsVIP1s and almost all OsPR1s were up-regulated and transformation efficiency of Y73 was increased notably when infected by Agrobacterium at 20 °C compared with 25 °C. This study suggests that the bacterial blight resistance genes in Y73 also have an effect on Agrobacterium-mediated transformation and that the response to temperature is associated with the regulation of MAPK/VIP1 defense signaling pathway.     

The effects of Artemisia annua L. and Achillea millefolium L. crude leaf extracts on the toxicity, development, feeding efficiency and chemical activities of small cabbage Pieris rapae L. (Lepidoptera: Pieridae)

The biological effects of two important medicinal plants, Artemisia annua L. and Achillea millefolium (L.) (viz, mortality, growth, and feeding indices as well as enzyme and non-enzymatic activities) were studied on small white Pieris rapae L a deleterious pest of cruciferous plants under controlled conditions (16:8 h L:D at 25 ± 1 °C and 65 ± 5% RH). The LC₅₀ and LC₂₅ values were 9.387% and 3.645% for A. annua L. and 4.19% and 1.69% for A. millefolium (L.), respectively. At the lowest concentration (0.625%), the deterrency was 29.826% and 44.185% for A. annua L. and A. millefolium (L.), respectively. Feeding indices were variously affected with changes in a number of parameters and an increase in larval and pupal duration. The activity level of alkaline phosphatase increased sharply while alanin and aspartate aminotransferases showed a sharp decrease. For non-enzymatic compounds, the amount of glucose and uric acid increased, but total protein and cholesterol decreased. These results indicate that these two medicinal plants might possess potential secondary metabolites that may be useful for controlling potential insect pests.     

枣镰翅小卷蛾成虫的寄主趋向和产卵选择

采用固相微萃取(SPME)和GC-MS分析研究木枣和酸枣挥发物成分,并测试枣镰翅小卷蛾触角电位(EAG)反应、寄主趋向和产卵选择,以探明枣镰翅小卷蛾的寄主选择机制。结果显示,在萌芽期,木枣和酸枣嫩叶的挥发成分均为罗勒烯、2-甲基-2-菠烯、α-法呢烯和邻苯二甲酸二丁酯4种,但相对含量稍有不同。枣镰翅小卷蛾成虫对木枣和酸枣2种寄主都有强烈的EAG反应,且同一寄主上雌蛾的EAG反应值极显著地高于雄蛾,其EAG值是雄蛾的3.3倍。枣镰翅小卷蛾对木枣表现强烈的趋向反应,而对酸枣的趋向不明显,且雌虫的趋向反应显著高于雄虫。木枣上的产卵量显著高于酸枣,且木枣枣吊上的单雌产卵量为307.9粒,极显著地高于酸枣枣吊上的产卵量(182.9粒)。研究表明,枣镰翅小卷蛾雌蛾在寄主选择中起主导作用,木枣是其嗜好寄主。     

Analysis of two acetylcholinesterase genes in Bombyx mori

Two acetylcholinesterases (AChE, EC 3.1.1.7) cDNAs were identified and cloned from silkworm, Bombyx mori. One of those, BmAChE-o cDNA, is comprised of 3197 nucleotides which encode 638 amino acids, having an amino acid sequence homology of 72% with Drosophila melanogaster Ace-orthologous AChE (AO-AChE). In some species, another AChE group based on the sequence, Drosophila Ace-paralogous AChE (AP-AChE) has been recognized in relation to organophosphate- or carbamate-resistance, but there have been few reports of AP-AChE among lepidopteran species. However, we isolated the AP-AChE from lepidopteran silkworm, and cloned full ORF as BmAChE-p, which cDNA consisted of 2465 nucleotides that encode 683 amino acids. The homologies with other AP-AChEs were over 60% when compared. Although silkworm is not a target of pesticides, the genomic information obtained in this study will contribute to insecticide-resistance study on lepidopteran pest species.     

菜豆普通细菌性疫病菌在土壤和植株残体中的越冬能力

为评估菜豆普通细菌性疫病菌地毯草黄单胞杆菌菜豆致病变种或褐色黄单胞菌褐色亚种在土壤及植物残体中的越冬能力,对采自黑龙江、内蒙古、山西、河北及新疆的18块菜豆生产田的20份土壤及14份植物残体样品进行病原菌分离和鉴定。在MT选择性培养基上有12个土壤样品和13个植株残体样品提取液产生典型的类似黄单胞菌菌落。选取29个分离物进行致病性测定,有27个分离物对菜豆品种"英国红"致病。利用地毯草黄单胞杆菌菜豆致病变种和褐色黄单胞菌褐色亚种的特异性引物X4c/X4e及褐色黄单胞菌褐色亚种特异性引物Xf1/Xf2对29个分离物进行多重PCR检测,其中17个分离物为地毯草黄单胞杆菌菜豆致病变种,10个分离物为褐色黄单胞菌褐色亚种。结果表明,菜豆普通细菌性疫病菌可以在黑龙江、内蒙古、山西、河北的一些菜豆种植区的土壤及植株残体中越冬存活。     

苏云金杆菌亚致死浓度对美国白蛾及其寄生蜂生长发育的影响

为阐明苏云金杆菌Bacillus thuringiensis(Bt)与白蛾周氏啮小蜂Chouioia cunea Yang的协同作用,分别用103~108孢子/m L的6个Bt浓度处理美国白蛾Hyphantria cunea 2龄幼虫后,通过室内试验测定了Bt对其致死、亚致死效应,及亚致死浓度Bt对寄生蜂子代出蜂的影响。结果表明,浓度106孢子/m L处理组美国白蛾均未能化蛹;而106孢子/m L处理组均能化蛹并羽化,且幼虫发育历期随Bt浓度的增加而延长,平均蛹重在74.92~92.18 mg之间,均高于对照的68.66 mg,但蛹重不呈线性规律变化。当Bt对美国白蛾平均校正死亡率低于17.17%时,幼虫发育历期延迟不明显,但平均蛹重为101.11~124.34 mg,均高于对照;接蜂后能够正常出蜂,出蜂率为33.13%~51.25%,每蛹出蜂量为117.43~143.42头,均显著高于对照。表明用平均校正死亡率低于20%的Bt处理对白蛾周氏啮小蜂的寄生有利,2种生防措施协同作用存在增效潜力。     

Biological and biochemical responses of Biomphalaria alexandrina to some extracts of the plants Solanum siniacum and Artemisia judaica L.

The molluscicidal activity of cold water, boiled water, methanol, ethanol, acetone and chloroform extracts of Solanum siniacum and Artemisia judaica L. plants against Biomphalaria alexandrina snails was carried out. The tests revealed plant’s ethanol extract was more toxic to the snails than the other tested extracts. Therefore, it was tested against snails’ fecundity (M_x), reproduction rate (R₀) and their infection with Schistosoma mansoni miracidia. In addition, biochemical parameters and the activities of some enzymes in tissues of snails treated with the two tested plants were determined. As well, glucose concentration in snails’ hemolymph was evaluated. Exposure of B. alexandrina snails to plant’s ethanol extract led to a significant reduction in their survival and snails’ fecundity, reproduction rate. In addition, it caused a considerable reduction in the infectivity of S. mansoni miracidia to the snails. Also, it caused a reduction in number of cercariae per snail during the patent period and in the period of cercarial shedding. The results revealed that the glucose concentration in hemolymph and Lactate level in soft tissues of treated snails were increased (P < 0.001) while glycogen, total protein, the lipid content and the pyruvate level in snail’s tissues decreased (P < 0.001). The activities of lactic dehydrogenase (LDH), pyruvatekinase (PK) and cytochrome oxidase (CY) enzymes in homogenate of snail’s tissues were reduced (P < 0.001) in response to treatment with the two tested plants while protease (PR) activity increased (P < 0.001). It is concluded that the application of LC₂₅ of ethanol extract of S. siniacum and A. judaica L. may be helpful in snail control as it interferes with the snails’ biology and physiology.     

两株棒束孢菌的鉴定及其对茶卷叶蛾和茶小卷叶蛾的致病力

为明确分离自茶卷叶蛾和中华大刀螳僵虫的2株虫生真菌的归属及生防潜力,采用形态学特征和rDNA-ITS序列分析其分类地位,测定了其对茶卷叶蛾和茶小卷叶蛾幼虫的致病力,并初步筛选了其最适培养条件.结果表明:2株真菌均为环链棒束孢Isaria cateniannulata,分别命名为ICBS918和ICTL911.在1.0×10⁸孢子/mL浓度下,ICBS918和ICTL911对茶卷叶蛾幼虫的累计校正死亡率均达100%,LT₅₀分别为3.13 d和3.15 d;对茶小卷叶蛾幼虫的累计校正死亡率分别为100%和95%,LT₅₀分别为3.25 d和3.31 d.接菌后8 d,2菌株对茶卷叶蛾幼虫的LC₅₀分别为0.47×10⁵孢子/mL和1.01×10⁵孢子/mL;对茶小卷叶蛾幼虫的LC₅₀分别为2.20×10⁵孢子/mL和1.34×10⁵孢子/mL.2菌株23 ℃下最适生长培养基均为综合马铃薯培养基,最适产孢培养基均为蛋白胨马铃薯葡萄糖琼脂培养基;在萨氏培养基上,菌株ICBS918和ICTL911分别在24~30 ℃及24 ℃时生长速度最快,在27 ℃和21 ℃时产孢量最大.表明2株环链棒束孢对茶卷叶蛾和茶小卷叶蛾幼虫均具有较好的生防潜力,可作为生防菌进行开发和应用.