Seed Transmission of Maize Dwarf Mosaic Virus in Sweet Corn

Sweet corn seed from several maize dwarf mosaic virus (MDMV)-infected hybrids grown in the field were tested for transmission of MDMV through the seed. Seeds collected in 1979, 1980, 1981, and 1982, were germinated in the greenhouse the following winters. Only one seedling of 22,189 was MDMV-infected During the last three years, seed were dissected at different maturities and the seed parts tested for the presence of MDMV by both infectivity and enzyme-linked immunosorbent assay (ELISA). At 21 days after pollination, MDMV always was detected in the pericarp, but rarely in the endosperm or embryo. No MDMV was detected in the embryo of mature kernels, but virus occasionally was detected in the endosperm and pericarp. MDMV was regularly detected in unfertilized kernels and whole silks, but not in pollen by infectivity, ELISA or serological specific electron microscopy. MDMV was detected in glumes and whole anthers.     

玉米矮花叶病毒研究进展

玉米矮花叶病毒 (Maizedwarfmosaicvirus,MDMV)是在世界范围内广泛分布的重要病毒 ,文中从寄主范围、传播、株系状况、分子生物学及检测等方面 ,对MDMV的国内外研究现状进行了阐述。     

玉米矮花叶病毒提纯及特性的研究

改进病毒纯化方法,获得了较高纯度、较强侵染活性的提纯病毒制品。提纯病毒的产量为0.7—1.2mg/100g病叶,病毒粒体大小为720—750×14nm,SDS-聚丙烯酰胺凝胶电泳测得的外壳蛋白分子量为36000道尔顿,用提纯病毒免疫家兔得到较高特异性的抗血清,微量沉淀反应的效价为1/2048,间接ELISA法测出的感病叶汁液的最高稀释倍数为3200—6400倍。     

RNA Interference-Based Transgenic Maize Resistant to Maize Dwarf Mosaic Virus

Maize dwarf mosaic virus (MDMV) is a widespread pathogenic virus that causes serious loss of yield in maize (Zea mays). RNA interference (RNAi) triggered by hairpin RNA (hpRNA) transcribed from a transgenic inverted-repeat sequence is an effective way to defend against viruses in plants. In this study, an hpRNA expression vector containing a sense arm and an antisense arm of 150 bp separated by an intron of the maize actin gene was constructed to target the P1 protein (protease) gene of MDMV and used to transform Agrobacterium tumefaciens strain EHA105. The transformed Agrobacterium strain was used to transform maize embryonic calli isolated from immature embryos by an improved culture technique. In all, 46 plants were regenerated after stringent hygromycin B selection, and 18 of them were certified to be positive by PCR amplification. Of these positive plants, 13 were grown to produce offspring, and nine were identified by Southern blotting to have the transgene integrated with one or two copies. The resistance of three T₂ lines was evaluated in a field trial of dual MDMV inoculation in two environments and was found to be improved compared with the non-transformed control. The disease indexes of the transgenic plant lines h2, 13, and h1 were not significantly different from the highly resistant control line H9-21. The viral titers of the inoculated plants were detected by double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), and the result was in accord with the resistance evaluated in the field trial. The addition of uniconazole S3307 (0.25 mg l⁻¹) and ABT root-promoting powder (0.5 mg l⁻¹) showed a significant improvement of hardening in regenerated plantlets, which were stronger and generated a better fibrous root system than the control. This improvement could facilitate the transgenic operation of maize.     

玉米矮花叶病研究进展

玉米矮花叶病(maize dwarf mosaic virus,MDMv)是世界上玉米产区普遍发生的病毒病害之一.自20世纪90年代以来,我国玉米矮花叶病发生严重,山西、甘肃、山东、河北以及北京等省市先后大流行,造成了巨大的农业经济损失.在我国玉米产区造成危害的主要是该病毒的B株系,主要借蚜虫传播和种子传播;在玉米矮花叶病的防治中,种植抗病品种,并辅以合理的栽培管理,可有效防止MDMV.本文主要综述玉米矮花叶病病毒的理化特性、玉米矮花叶病的发生危害、病原及其传播方式、发病条件、流行与防治、品种(自交系)抗性、抗性鉴定、抗性遗传及其抗病基因工程研究等方面的研究进展,以期为以后玉米矮花叶病的有效防治提供一定的参考.     

杭州地区发生的玉米花叶病由甘蔗花叶病毒引起

从杭州地区呈现玉米矮花叶典型症状的玉米病组织中提纯得到大量线状病毒粒子,大多数长度为750nm。病组织中含有大量风轮状内含体和板状集结体,病毒外壳蛋白为33.6kD。病毒RNA1 3'端序列（1.8kb）与甘蔗茶花经叶病毒（SCMV）同源性最高,达71.5%-99.1%,与高梁花叶病毒（SrMV）同源性次之,为67.8%-68.5%,与玉米矮花叶病毒（MDMV）同泊性最低,仅为38.4%-48.4%,从而初步认为此病害由SCMV引起。根据已发表的SCMV外壳蛋白氨基酸序列作亲缘性分析,表明SCMV可分为美国、南非、澳大利亚、德国和中国三大类。     

杭州地区发生的玉米花叶病由甘蔗花叶病毒引起

从杭州地区呈现玉米矮花叶典型症状的玉米病组织中提纯得到大量线状病毒粒子,大多数长度为750?nm。病组织中含有大量风轮状内含体和板状集结体。病毒外壳蛋白为33.6 kD。病毒RNA13’端序列(1.8 kb)与甘蔗花叶病毒(SCMV)同源性最高,达71.5%～99.1%,与高梁花叶病毒(SrMV)同源性次之,为67.8%～68.5%,与玉米矮花叶病毒(MDMV)同源性最低,仅为38.4%～48.4%,从而初步认为此病害由SCMV引起。根据已发表的SCMV外壳蛋白氨基酸序列作亲缘性分析,表明SCMV可分为美国、南非、澳大利亚;德国和中国三大类。     

MDMV CP基因的克隆及其转基因玉米的研究

用RT-PCR方法分离了玉米矮花叶病毒外壳蛋白基因(MDMV CP),并且利用基因枪法将该基因导入玉米优良自交系18-599红、18-599白幼胚诱导的愈伤组织中。转化的愈伤组织在Bialaphos浓度(PPT)为8mg／L、10mg/L、5mg／L的筛选压下经过3次抗性筛选后,分别再生出可育植株12株和6株。PCR和Southem检测结果说明CP基因已整合到玉米自交系基因组中。对T1代转基因植株进行病毒人工接种试验,结果表明对照植株全部表现为感染玉米矮花叶病的典型症状,而转基因植株后代呈现不同程度的抗性。     

转基因抗矮花叶病玉米的遗传、表达及抗病性研究

目的：研究目的基因在转基因植株及其后代中遗传表达的稳定性,以及目的基因表达与抗病性的关系,最终得到转基因纯合株系。方法：以采用花粉介导法将RDV运动蛋白缺陷型（RDV MP^-）基因导入玉米自交系478的转基因种子（T0）作为试验材料,对其或其后代进行潮霉素抗性筛选、PCR检测、目的基因表达产物含量测定、农艺性状筛选,以及田间接种病毒的抗病鉴定。结果：通过潮霉素抗性筛选从T0种子获得了11株疑似转化植株;对T1、T2、T3代转基因植株的PCR分析证实目的基因已导入玉米植株,并显示随着转化植株世代交替,目的基因可稳定遗传给下一代,且目的基因在待测材料中的检出比例也随着代数的增加而提高;目的基因表达量的测定结果为1．83-11．57ng／mg叶片鲜重之间;田间接种玉米矮花叶病病毒试验结果证明转化植株比对照植株的抗矮花叶病能力有了显著提高,个别株系在T1代的发病率就为0,T1、T2、B代转化植株的抗病性逐代提高,比临近对照的抗病性提高2～5级;目的基因表达量与植株（系）的抗病性显著相关,r=0．923,P〈0．01;入选纯合系的农艺性状也有较大变化,穗粒数比对照系增加约5％。结论：通过以上方法,可以筛选到转基因抗病玉米纯合株系。     

The Purification and Characterization of Gemini Particles from Abutilon Mosaic Virus Infected Malvaceae

Gemini particles were purified from Abutilon Mosaic Virus infected Abutilon sellovianum, Sida micrantha and Malva parviflora. They sediment with 82 S, possess a density of 1.30 g/cm³ in Cs₂SO₄ and a nucleic acid percentage of 18 % calculated from ultra-violett spectrometry. Their proteins are separated in SDS-PAGE into two components of 28 000 and 27 000 mol. wt. Their nucleic acid consists of single-stranded circular DNA with length distribution maxima of 2 800, 2 500 and 1 300 bases for the circular molecules and 2 050, 1 850, and 1 300 bases for the correspondmg linear molecules.     

烟草花叶病毒对烟草叶片光合特征和POD表达的影响

以烤烟(Nicotiana tabacum L.)品种'中烟5号'为实验材料,对烟草健康株与感染烟草花叶病毒(TMV)株的叶绿素、光合速率、光合速率对光强的响应曲线、光暗反应荧光特征、POD活性及其表达等进行研究,以探讨TMV感染对烟草植株生理生态特征的影响.结果显示:病株的叶绿素a(Chl a)和叶绿素b(Chl b)含量显著低于健康株,但Chl a/Chl b值基本相同;病株暗中初始荧光(F0)、暗中最大荧光(Fm)、暗中可变荧光(Fv)、光下初始荧光(F0′)、光下最大荧光(Fm′)、光下可变荧光(Fv′)、非光化学猝灭系数(NPQ)、PSⅡ捕光效率(Fv′/Fm′)、PSⅡ实际光化学效率(ФPSⅡ)及光饱和点显著低于健康株;净光合速率在光强较大(＞1 500 μmol·m-2·s-1)时病株比健康株低,光强适中(1 500 μmol·m-2·s-1左右)时两者相差不大,光强较弱(＜1 500 μmol·m-2·s-1左右)时病株比健康株高;病株叶片的过氧化物酶(POD)活性显著升高,POD同工酶中一些大分子量蛋白分子表达量加大.研究表明,感染TMV使烟草植株对光抑制更为敏感,叶片的荧光激发能力和热耗散能力下降,PSⅡ反应中心捕光效率和光化学反应效率降低,光合电子传递能力和碳同化能力受到抑制;POD活性提高和表达量增加可能是诱导烟草抗病性的一个关键生理过程.     

两个玉米矮花叶病显性互补抗病基因的发现和定位

玉米矮花叶病是世界普通发生危害严重的玉米病毒病害之一,迄今为止,只有少数几个抗病基因被发现并定位,优良自交系四一是鉴定出定的玉米筹花叶病新抗源,它表现为全生育抗性,通过连续两年的经典遗传学研究发现,四一的成株期抗性表现为一种新的抗病遗传模式,该抗性是由两个显性互补抗病基因控制,87对微卫星标记分析进一步证实了以上推论,并把两个抗病基因分别定位在第三和第六染色体上,第三染色体上的抗病基因与微卫星标记phi029相距14.5cM,第六染色体上的抗病基因与微卫星标记phil26相距7.2cM.     

玉米种质资源抗矮花叶病鉴定

采用摩擦接种方法,鉴定了3995份玉米种质对玉米矮花叶病毒(SCMV-MDB)的抗病性。经重复鉴定,筛选出抗病优良自交系73份,抗病丰产杂交种80份。玉米不同类群种质,对SCMV-MDB的抗病性有明显差异,改良Reid、Lancaster、旅大红骨类群种质大部分表现感病;塘四平头、外杂先锋选系类群种质大多数表现抗病。在不同类型种质中,硬粒型、马齿型和半马齿型表现抗病的较多,糯质型较少,甜质型和爆裂型中尚未发现抗源。     

Location of Grapevine Fardeaf and Yellow Mosaic Virus Particles in Xiphinema index

Particles of fanleaf and yellow mosaic viruses are reported in the lumen of the esophagus of Xiphinerna index. Differences in cuticular morphology suggest differences in charged receptor sites which may offer an explanation for virus location and orderly arrangement.     

玉米矮花叶病毒CP基因dsRNA的原核表达与分离

根据玉米矮花叶病毒CP基因序列设计特异性引物,RT-PCR扩增玉米矮花叶病毒CP基因特异性干涉片段,将干涉片段及pUCCRNAi载体分别用BamH I及Sal I双酶切,然后将干涉片段分别正反向插入pUC- CRNAi载体中,构建CP基因反向重复克隆载体pUCCRNAi+2 F.再利用Pst I-Sal I位点插入到L...     

Effect of Source Strength, Distance and Direction on the Spread of Maize Dwarf Mosaic Virus

Transmission of Pseudomonas avenae from rice seed to seedling and from plant to seed was shown. Based on experiments with unhulled and hulled seeds and on histopathological studies the location of the pathogen in seeds and the possible pathway of the pathogen from seed to seedling is suggested. Infected seeds were harvested from plants on which no symptoms were observed after the seedling stage. The experimental conditions indicated that the bacterium can be transmitted internally from plantto seed in latently infected plants.