侵染菠菜的芜菁花叶病毒鉴定

菠菜病毒分离物Ta-1可侵染甘蓝等8科25种植物,但不侵染心叶烟。病汁液和芜菁花叶病毒抗血清呈阳性反应。光学显微镜下观察到受病毒侵染的细胞内有不定形内含体。从超薄切片见到细胞质中有风轮状内含体存在。电镜观察其粒子形态为均一的线状病毒,大小为700—760×10—12.5nm。部分提纯病毒紫外吸收呈典型核蛋白吸收曲线。SDS—PAGE法测得病毒外壳蛋白亚基分子量为35,500。     

萝卜优异种质对芜菁花叶病毒抗性的遗传分析

为了深入认识萝卜优异种质对芜菁花叶病毒(TuMV)的抗性遗传机制,本研究以高抗TuMV的自交系Q07-12(P_1)和高感自交系KB07-10(P_2)为亲本,构建了包括P_1、P_2、F_1、B_1、B_2及F_2的6个群体,采用数量性状主基因+多基因混合遗传的六世代联合分析法,对其抗性遗传规律进行了研究。结果表明,萝卜对TuMV的抗性受两对主基因控制,并符合"加性-显性-上位性主基因+加性-显性-上位性多基因"模型,其中主基因遗传率在B_1、B_2及F_2群体中分别为89%、55%和95%,多基因的遗传率分别为0、40.9%和0,环境因素对抗性强弱的影响较小。此结果表明了该优异种质在萝卜抗病毒遗传改良中的利用价值,为萝卜抗病育种和抗病基因的挖掘提供了理论依据。     

芜菁花叶病毒(TuMV)特性的研究进展

芜菁花叶病毒(TuMV)是世界范围内广泛分布的重要病毒。从生物学和理化特性、基因组结构、侵染和繁殖、变异及利用转基因技术提高病毒抗性等方面对TuMV的国内外研究现状进行了阐述。     

芜菁花叶病毒分子生物学研究进展

芜菁花叶病毒(Turnip mosaic virus,TuMV)是马铃薯Y病毒属(Potyvirus)的一个重要成员, 在世界上分布广泛,主要危害十字花科植物。本文对芜菁花叶病毒的分子生物学相关研究,如基因组结构和功能、株系变异与进化、寄主植物抗性以及抗病毒转基因工程等的研究进展作一阐述。     

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。     

玉米种质资源对六种重要病虫害的抗性鉴定与评价

在2003-2005年间,对604份玉米种质进行了抗弯孢菌叶斑病和玉米螟鉴定,筛选出抗弯孢菌叶斑病的材料93份,抗玉米螟材料22份。2006-2009年间,对836份玉米种质进行了抗大斑病、茎腐病、穗腐病和瘤黑粉病的鉴定与评价,筛选出一批高抗和多抗的资源。在836份资源中,对大斑病1、2和N号3个生理小种具有抗性的材料均为50%左右;抗茎腐病材料为41.3%,高抗和抗性种质分别为264和81份;穗腐病高抗和抗性种质分别为5和171份,占比为21.1%;瘤黑粉病高抗和抗性种质各261和14份,占总鉴定材料的32.9%。上述结果表明抗大斑病、茎腐病和瘤黑粉病的种质资源较为丰富。通过对抗性结果进行对比分析,发现不同生态区玉米种质的抗性强弱以及抗性多样性存在明显差异,黑龙江和内蒙古的种质对病虫害的抗性强弱及多样性程度明显高于四川种质。此外,玉米自交系对病虫害的抗性强弱以及多抗性程度高于农家种。     

玉米种质资源对六种重要病虫害的抗性鉴定与评价

在2003－2005年间,对604份玉米种质进行了抗弯孢菌叶斑病和玉米螟鉴定,筛选出抗弯孢菌叶斑病的材料93份,抗玉米螟材料22份。2006－2009年间,对836份玉米种质进行了抗大斑病、茎腐病、穗腐病和瘤黑粉病的鉴定与评价,筛选出一批高抗和多抗的资源。在836份资源中,对大斑病1、2和N号3个生理小种具有抗性的材料均为50％左右;抗茎腐病材料为41.3％,高抗和抗性种质分别为264和81份;穗腐病高抗和抗性种质分别为5和171份,占比为21.1％;瘤黑粉病高抗和抗性种质各261和14份,占总鉴定材料的32.9％。上述结果表明抗大斑病、茎腐病和瘤黑粉病的种质资源较为丰富。通过对抗性结果进行对比分析,发现不同生态区玉米种质的抗性强弱以及抗性多样性存在明显差异,黑龙江和内蒙的种质对病虫害的抗性强弱及多样性程度明显高于四川种质。此外,玉米自交系对病虫害的抗性强弱以及多抗性程度高于农家种。     

玉米种质对黄曲霉菌抗性的鉴定

以145个玉米自交系作为试验材料,对其籽粒人工接种黄曲霉菌（Aspergillus flavus),根据籽粒的发病情况进行抗性鉴定。结果表明：（1）各玉米自交系对黄曲霉菌的抗性有极显著的差异,在145个自交系中,高抗的8份,中抗的72份,中感的57份,高感的8份;（2）对黄曲霉菌有较高抗性的自交系大多数来自于我国的南方地区;（3）通过辐射处理,对于提高抗性可能有较好的效果;（4）爆裂玉米具有较好的抗性;（5）不同致病菌株接种,抗性反应有差异。     

芜菁花叶病毒单克隆抗体的制备及检测应用

先以芜菁花叶病毒（TuMV）免疫BAL B/C小鼠,然后取其脾细胞使之与SP2/0鼠骨髓瘤细胞融合,经筛选、克隆,获得4株能稳定传代并分泌抗TuMV单克隆抗体（Mab）的杂交瘤细胞,并以之制备腹水单抗。4株单克隆抗体腹水ELISA效价在10-5～10-6之间,仅对TuMV起特异性反应。Western blot分析表明,4株单抗都能与TuMV 34kD的外壳蛋白亚基起特异反应。利用TuMV的多抗兔血清和单抗腹水建立了三抗体夹心ELISA检测TuMV的方法,检测病叶的灵敏度为1∶5120倍,检测提纯TuMV病毒绝对量为21.9 ng。利用三抗体夹心ELISA测定出7种作物上有TuMV侵染。      

马铃薯种质资源的晚疫病抗性鉴定

采用离体叶片接种法,对43份马铃薯种质资源进行晚疫病抗性鉴定、比较和评价。以接种5 d后的感病品种‘Désirée’和抗病品种‘加湘1号’叶片症状为对照,鉴定出5份表现为高抗的种质资源,其中包含3个Solanum phurejia和2个S. tuberosum ssp. andigena材料。另外,还鉴定出14份中抗材料(No. 7～20)。结果表明,野生种和安第斯山栽培亚种马铃薯资源的抗性材料较为丰富,可作为晚疫病抗病育种的亲本。     

Draft Sequences of the Radish (Raphanus sativus L.) Genome

Radish (Raphanus sativus L., n = 9) is one of the major vegetables in Asia. Since the genomes of Brassica and related species including radish underwent genome rearrangement, it is quite difficult to perform functional analysis based on the reported genomic sequence of Brassica rapa. Therefore, we performed genome sequencing of radish. Short reads of genomic sequences of 191.1 Gb were obtained by next-generation sequencing (NGS) for a radish inbred line, and 76,592 scaffolds of ≥300 bp were constructed along with the bacterial artificial chromosome-end sequences. Finally, the whole draft genomic sequence of 402 Mb spanning 75.9% of the estimated genomic size and containing 61,572 predicted genes was obtained. Subsequently, 221 single nucleotide polymorphism markers and 768 PCR-RFLP markers were used together with the 746 markers produced in our previous study for the construction of a linkage map. The map was combined further with another radish linkage map constructed mainly with expressed sequence tag-simple sequence repeat markers into a high-density integrated map of 1,166 cM with 2,553 DNA markers. A total of 1,345 scaffolds were assigned to the linkage map, spanning 116.0 Mb. Bulked PCR products amplified by 2,880 primer pairs were sequenced by NGS, and SNPs in eight inbred lines were identified.     

春萝卜抽薹过程中内源激素含量变化分析

采用酶联免疫吸附法(ELISA)测定了春萝卜抽薹过程中顶端生长点处嫩叶的赤霉素(GA₄)、生长素(IAA)、细胞分裂素(iPA)和脱落酸(ABA)等含量的变化情况。研究结果表明,在萝卜抽薹过程中,GA₄含量在现蕾点存在明显的峰值,表明GA₄与现蕾抽薹密切相关,起着主导作用;内源ABA含量变化与GA₄变化趋势基本一致;与GA₄和ABA含量相比,抽薹过程中叶片IAA含量相对较低;iPA含量在现蕾期逐步增加,至抽薹中后期出现一高峰值。研究发现iPA/ABA值增大,GA₄/iPA和GA₄/IAA值变小有利于花芽分化后萝卜植株抽薹开花,表明内源激素之间的平衡对萝卜抽薹开花也起着重要作用。     

不结球白菜种质资源对TuMV的抗性鉴定与评价

分别利用苗期人工接种鉴定及ELISA检测方法,对127份不结球白菜种质资源进行芜菁花叶病毒(TuMV)的抗性鉴定。结果显示,苗期人工接种鉴定的病情指数(DI)分布在3.55~95.68之间,不同种质间表现出较大的抗性差异。不同抗性级别的次数分布图基本符合正态分布,略向感病区域偏离。基于病情指数的聚类分析结果与抗病性分级基本一致。通过苗期鉴定共筛选获得高抗TuMV的不结球白菜种质6份,抗病种质13份,其主要农艺性状表现出一定的多样性。其中叶面皱缩和具有刺毛的抗病材料所占比例较高,可能与TuMV抗性存在一定的相关性。ELISA检测结果显示,117份供试种质的P/N值分布在3.10~25.37之间,不同种质间表现出病毒含量的差异,但与DI值未呈现明显相关性,可作为抗病材料筛选的辅助指标。     

Genetic Diversity of Radish (Raphanus sativus L.) Germplasm Resources Revealed by AFLP and RAPD Markers

Genetic diversity of 56 radish accessions, representing nearly all the typical types and origins of cultivated radish germplasms conserved in the National Mid-term Genebank for Vegetables of China, was assessed with amplified fragment length polymorphism (AFLP) and random amplified polymorphic DNA (RAPD) markers. A total of 72 and 128 polymorphic bands were generated by the 12 selected RAPD primers and eight AFLP primer combinations respectively. A moderate correlation with the value of r = 0.66 was observed between AFLP and RAPD markers. The total 200 polymorphic bands were integrated to assess the genetic diversity of 56 radish accessions. The Jaccard similarity coefficients between the accessions varied from 0.30 to 0.83 with the mean of 0.54. Cluster analysis classified the germplasms into three groups of var. hortensis Becker, var. sativus, and var. niger Kerner. The three-dimensions scatter plot of principle coordinate analysis (PCA) further divided var. hortensis Becker germplasms into two separate groups. The results indicated that the genetic diversity harbored among var. hortensis Becker germplasms was very abundant, which could be further exploited for radish genetic improvement.     

萝卜种质资源对黑腐病的抗性鉴定与筛选

对40份初选萝卜种质分别接种Xcc8004和XccBJ两个菌株,进行黑腐病苗期抗性鉴定,对其中8份代表性萝卜种质肉质根切片接种Xcc8004进行抗性鉴定和27份萝卜种质幼苗接种8个效应物基因进行过敏反应鉴定。结果表明:不同萝卜种质苗期对黑腐病的抗性存在显著差异,筛选出高抗Xcc8004的材料3份、抗病1份、中抗4份,高抗XccBJ的材料1份、抗病2份、中抗5份。萝卜苗期对Xcc8004和XccBJ的抗病性极显著相关,幼苗和肉质根对Xcc8004的抗病性极显著相关。筛选出17份对不同效应物表现过敏反应的萝卜种质。对效应物XC₀241表现过敏反应的种质数最多,对XC₀542和XC₀541表现过敏反应的种质数次之。不同抗源对不同效应物的过敏反应程度有所不同。稳定可靠抗病资源的获得为萝卜抗病育种和抗病机理的深入研究提供了基础材料。     

Natural Infection of Field-Grown Daikon Radish (Raphanus sativus L.) Crops by Turnip Crinkle Virus in North America

In 1993, stunted field-grown daikon radish plants were found with black lesions on the petioles and leaf blades, and new growth exhibited leaf crinkling and a mosaic pattern on distorted leaves. The plants had hollow roots and seed production was severely reduced. Symptomatic plants contained isometric virions identified as those of turnip crinkle virus (TCV). This is the first report of TCV causing serious economic damage and the first report of natural infection in North America.     

Heavy Metal Phytotoxicity to Radish (Raphanus sativus L.) in a Digested Sludge-amended Gangetic Alluvium

A limiting factor in land application of sewage sludge is the resultant heavy metal accumulation in soils followed by biomagnification in the food chain, posing a potential hazard to animal and human health. In view of this fact, pot experiments were conducted to evaluate the effect of digested sludge application to soil on phytotoxicity of heavy metals such as Cd, Cr, Ni, and Pb to radish (Raphanus sativus L.) plants. Increasing sludge levels resulted in increased levels of DTPA-extractable heavy metals in the soil. Cadmium was the dominant metal extracted by DTPA followed by Ni, Pb, and Cr. The extractability of metals by DTPA tended to decrease from the first to the second crop. Dry matter yield of radish increased significantly as a function of increasing sludge treatments. Soil application of sludge raised the concentration of one or more heavy metals in plants. Shoots contained higher concentrations of Cd, Cr, and Ni than the roots of radish plants. Shoot concentrations of Cd, Cr, Ni, and Pb were within the tolerance levels of this crop at all rates of sludge application. Shoot as well as root concentration of Cd was above 0.5 mg kg^?1, considered toxic for human and animal consumption. The levels of DTPA-extractable Cd and Ni were less correlated while those of Cr and Pb were more correlated with their respective shoot and root contents. The results emphasize that accumulation of potentially toxic heavy metals in soil and their build-up in vegetable crops should not be ignored when sludge is applied as an amendment to land.     

Subcellular Localization and Possible Functions of γ-Glutamyltransferase in the Radish (Raphanus sativus L.) Plant

Previously we reported the purification of soluble γ-glutamyltransferases (GGTs) from radish cotyledon. Subcellular fractionation of radish cells revealed that soluble GGT is a vacuolar enzyme. Acivicin, a GGT inhibitor, mediated the in vivo catabolism inhibition of the glutathione S-conjugate generated from endogenous glutathione and exogenously supplied monochlorobimane. Thus soluble GGT is possibly involved in the catabolism of glutathione S-conjugates.     

萝卜颗粒引发大豆种子抗吸胀冷害能力的效应

以鲜萝卜颗粒和聚乙二醇(PEG)引发大豆种子抗吸胀冷害能力的研究结果表明,二者都能引发和控制种子在低温吸胀过程中的吸水,提高种子活力和萌发率,减少细胞中电解质和氨基酸外渗。