茄子种质资源青枯病抗性鉴定及分子标记研究

利用分子标记结合人工接种鉴定评价41份茄子材料的青枯病抗性。试验结果表明,41个材料中,人工接种鉴定为抗病材料有38份,感病的3份,与分子标记鉴定结果基本吻合。综合人工接种与分子鉴定结果得出,只含有一个特异标记或不含特异标记的材料表现为感病。在3个分子标记鉴定结果中,有8份材料不完全一致,原因可能是各标记所连锁的抗性位点不同;另外,不同的材料在试验环境中基因表达很可能存在差异。因此,在茄子抗青枯病育种中,人工接种鉴定和分子标记辅助鉴定都是重要的。     

辣椒种质资源抗青枯病的鉴定与评价

采用青枯菌FJC100301菌株对田间辣椒（Capsicum annuum）抗病品种76a和感病品种TW-1分别作了不同温度、不同接种量和不同接种方法的接种试验。结果表明,辣椒青枯病抗性的室内鉴定以接种温度28℃、浸根20 min和3×10^8cfu/mL接种浓度为宜;辣椒种质田间抗青枯病接种鉴定宜选择5月上旬进行,浸根20 min,接种浓度为3×10^8cfu/mL。采用田间抗性接种鉴定的方法,用青枯菌FJC100301菌株对106份辣椒材料进行了抗性鉴定。田间接种后每隔10 d统计病情指数,划分辣椒抗青枯病鉴定分级标准,获得了高抗材料14份、抗病材料8份、中抗材料23份、中感材料23份、感病材料20份、高感材料18份;采用离体叶片接种法对田间筛选得到的高抗和高感纯度较高品种进行抗性分析,结果与田间鉴定一致。     

茄子栽培种砧用种质农业生物学性状及嫁接适用性研究

青枯病是热带亚热带地区普遍发生的一种土传病害,为选配具有优良抗病性和嫁接适用性的茄子砧木杂交组合,该研究对国内外11份茄子栽培种砧用种质进行农业生物学评价和青枯病抗病性鉴定,研究了不同砧用种质对嫁接苗生长的影响。结果表明:供试砧用种质中,果型为卵圆形5份、倒卵形2份、长卵圆4份;幼果颜色紫色7份、淡紫色1份、绿色1份、淡绿色2份;老熟果实颜色均为黄色;种质J14、J15、J16、J17的主茎紫色,其余均为绿色;J61的花瓣白色,其余均为紫色或淡紫色。供试砧木种质与接穗Rf的嫁接成活率均在80%以上,最高达97%。采用苗期伤根-浸根接种法鉴定砧木种质对青枯病的抗病性,有9份砧用种质自根苗表现高抗(HR),病情指数为3.67~12.33,其中6份砧木种质(BC01、BC03、BC04、BC06、J15、J16)嫁接感病接穗后表现抗病(R),病情指数为16.67~21.00。砧用种质能显著改善接穗的生长,其中,BC01、BC03、BC04、BC06、J15、J16的嫁接苗的生物量增长率显著高于其他种质的嫁接苗。综合试验结果显示茄子栽培种砧用种质农业生物学性状表现多样性,可区分为不同生态类型。抗病砧木种质嫁接高感病接穗的防病效果显著,种质BC01、BC03、BC04、BC06、J15、J16高抗青枯病,嫁接高感病品种表现抗青枯病,可作为配制抗青枯病砧木杂交组合的候选亲本。     

长江流域棉区棉花新品种抗病虫性研究

2003年,对长江流域棉区15份新材料进行了抗红铃虫鉴定,结果有8份材料高抗红铃虫,占鉴定材料的53.33%;4份材料抗红铃虫,占26.67%;3份材料中抗红铃虫,占20.00%.对55份新材料进行抗枯萎病鉴定,结果有27份材料抗枯萎病,占鉴定材料的49.09%;26份材料耐枯萎病,占47.27%;2份感枯萎病,占3.64%.对59份材料进行抗黄萎病鉴定,结果有8份材料抗黄萎病,占鉴定材料的13.56%;43份材料耐黄萎病,占72.88%;8份材料感黄萎病,占13.56%.     

云南野生茄资源黄萎病苗期人工接种抗性鉴定分析

黄萎病是目前茄子生产中的主要病害之一,广泛收集、鉴定、筛选抗性资源,尤其是从野生近缘种中发掘抗性基因并培育抗病品种,是解决茄子黄萎病危害的优选途径。云南省拥有丰富的野生茄子资源但尚未有效利用。本研究针对云南省茄子主产区收集到的3种黄萎病菌株,通过形态学鉴定和真菌18S r DNA/ITS鉴定,均属于大丽轮枝菌,并通过致病力鉴定,筛选出一株强致病力菌株(QZ-S);应用菌株QZ-S,通过苗期人工接种的方法对45份云南野生茄子资源开展黄萎病抗性鉴定,最终筛选出2份高抗材料(蒜芥茄和喀西茄)、2份抗病材料(水茄和多裂水茄)、6份中抗材料(1份刺天茄和5份红茄);此外,还筛选到了1份黄萎病高感材料239-3-2。本研究筛选出的材料可应用于茄子黄萎病抗病育种,为茄子及其他作物黄萎病抗病育种提供抗源。     

我国蚕豆品种资源对蚕豆锈病的抗性鉴定

1998～2000年对241份蚕豆品种(系)进行抗蚕豆锈病鉴定,其中表现高抗(HR)的材料2份,占0.83%;抗病(R)和中抗(MR)的材料131份,占54.36%.中感(MS)和感病(S)材料104份,占43.15%,高感(HS)的材料4份,占1.66%.结果表明,不同地理来源、株高、单株粒数、单株产量及粒重的材料抗性有差异.     

高温胁迫下华南茄子的耐热性表现及其鉴定指标的筛选

以6份茄子为材料研究了高温胁迫对幼苗热害指数、细胞膜相对电导率、脯氨酸含量的影响及其与田间自然鉴定结果的对应关系,结果表明,分段高温处理的热害指数和连续高温处理下的细胞膜电导率可以有效区分不同茄子材料幼苗的耐热性,且两者鉴定结果与田间鉴定结果基本一致,可作为茄子苗期室内快速鉴定的首选指标;脯氨酸含量变化没有规律,较难反映不同茄子幼苗间的耐热性差异;此外,对青枯病的抗性可以作为南方茄子耐热性综合评价的辅助指标.春季和秋季茄子开花、结果的比较研究表明,高温胁迫下坐果率、商品果率、果长、单果重和商品果产量有较大幅度下降,但不一定造成茄子正常花率的降低,果周径所受影响也较小.     

玉米重要自交系的肿囊腐霉茎腐病抗性鉴定与评价

由肿囊腐霉菌(Pythium inflatum Matthews)引起的玉米茎腐病是影响玉米产量的一种重要病害。为进一步拓展可利用的抗源,于2010-2011年在田间采用人工接种方法对287份重要的玉米自交系种质进行了玉米茎腐病的抗性鉴定评价。结果表明,287份鉴定材料中有171份自交系对茎腐病的抗性达到中抗以上水平,占鉴定材料的59.58%,其中高抗自交系共43份,占鉴定材料总数的14.98%;感病类型自交系共116份,占鉴定材料的40.42%,其中高感自交系共95份,占鉴定材料总数的33.10%。Lancaster、Reid及P群种质中具有丰富的茎腐病抗源,而塘四平头种质群中茎腐病抗源相对缺乏,多为感病类型。该研究结果可为今后我国玉米茎腐病抗性种质的引进和改良提供重要参考。     

抗青枯病烟草种质资源在云南省的评价

筛选出抗性稳定的种质资源是选育抗病品种的重要基础。本文采用人工接种和病田自然发病方法鉴定了48份烟草种质的青枯病抗性表现。土壤盆栽接种鉴定表现为高抗的材料有CF207、岩烟97、TI448A、DB101、G80、RG17、GTH-1等7份材料,表现抗病的有MSK149、Oxford 2028、NC95、YN108、K346、K358、Enshu FC、Oxford 207、RG11等9份材料。苗期恒温水培接种鉴定结果表明,Oxford 207和岩烟97表现为高抗,Enshu FC表现抗病,抗病材料与云烟85和K326杂交F1的抗性表现为中感至抗病。田间自然发病鉴定结果表明,我国审定的中抗青枯病的品种RG17、RG11、K358和K346,在云南省田间抗性表现为中抗,产值较高。TI448A田间表现为抗青枯病,但易感黑胫病和空茎病。Oxford 2028和Oxford 207田间表现为抗病至高抗,产值较高。G3和岩烟97田间分别表现为高抗和抗病,产值较低。根据接种鉴定和田间病圃2年抗性鉴定,筛选出育种潜力较大的青枯病抗源Oxford 207、Enshu FC,岩烟97和TI448A。     

广西玉米种质资源对纹枯病的抗性鉴定

在人工接种条件下,对860份玉米材料进行了纹枯病抗性鉴定和评价,旨在为进一步开展玉米纹枯病抗性育种和分子生物学研究奠定基础。结果显示,在鉴定的860份玉米材料中,没有发现免疫的材料,高抗、抗、中抗、感和高感的比例分别为3.49%、28.60%、26.16%、10.35%和31.40%;在农家种、群体种和杂交种（组合）等杂合体中,抗病材料所占比例较大,而自交系等纯合体中,感病材料所占比例较大,表明玉米杂合体种质资源材料中可能蕴藏着不同的抗纹枯病基因,特别是广西农家种中可能存在对纹枯病具有稳定抗性的材料,值得进一步研究和利用。     

花生抗青枯病种质脂肪酸组成的遗传多样性

通过对123份不同类型抗青枯病花生种质种子脂肪酸的鉴定测试,分析了抗青枯病花生种质在这些性状方面的遗传分化,并与6006份资源组成的花生基础收集品进行了比较。研究结果表明,我国抗青枯病花生资源的油酸含量平均为51.78%,显著高于基础收集品的对应值(45.64%);亚油酸含量平均为28.88%,显著低于基础收集品的对应值(34.36%);高油酸种质较多,油酸含量达61%以上的资源23份,所占比重为18.7%,显著高于基础品中的相应比重(2.65%)。标准差、变异系数以及遗传多样性指数的分析结果表明,抗青枯病资源在油酸和亚油酸含量方面的遗传分化程度高。     

黄萎病菌胁迫下野生茄子托鲁巴姆防卫反应的生理生化分析

以野生茄子托鲁巴姆(Solanum torvum Swartz)和苏崎茄(Solanum melongena L.)为主要材料,通过比较两种茄子在大丽轮枝菌(Verticillium dahliae Kleb)侵染过程中体内的生理生化指标变化,分析托鲁巴姆对黄萎病的抗性响应机制。结果表明:(1)与苏崎茄植株相比,托鲁巴姆植株表现出很强的自我防御和自我修复能力。(2)托鲁巴姆体内存在的活性氧清除系统(如SOD、POD、CAT等酶的活性)高于苏崎茄;在侵染后,托鲁巴姆体内各种酶的活性快速增加,其幅度高于苏崎茄;MDA的变化则恰恰相反。这个结果提示,黄萎病菌胁迫可能激活了托鲁巴姆体内活性氧清除系统,从而加快了某些防御物质(如木质素和抗菌物质绿原酸等)的形成,同时减缓或降低MDA等有害物质在植株体内的积累。(3)黄萎病菌侵染后各生理指标的响应时间表现出差异。托鲁巴姆中POD、PAL的活性和可溶性蛋白含量在侵染后的12h内就迅速作出响应(POD增加、PAL和可溶性蛋白减少);而SOD、PPO、CAT的活性和MDA的含量则在处理后初始阶段(至少12h)进行了一些调整,随后才进入持续性的增加或减少阶段。由此可见,托鲁巴姆对黄萎病病菌侵染的响应具有时序性,其体内POD酶、PAL酶和可溶性蛋白首先参与植物的防卫反应以应对黄萎病的胁迫,其它酶随后参与响应,它们的共同作用形成对黄萎病菌的有效防卫。     

Early burst of reactive oxygen species positively regulates resistance of eggplant against bacterial wilt

Reactive oxygen species (ROS) play a crucial role in the early response to plant biotic and abiotic stresses. In this study, bacterial wilt‐resistant and wilt‐susceptible eggplants were inoculated with Ralstonia solanacearum and the ROS content was analysed. The result revealed an increased accumulation of hydrogen peroxide (H₂O₂) and superoxide (O₂^?) in resistant and susceptible eggplant roots after R. solanacearum inoculation. H₂O₂ and O₂^? accumulation increased earlier in the inoculated resistant eggplant root than in the inoculated susceptible eggplant root. Real‐time polymerase chain reaction results revealed that respiratory burst oxidase homologue (Rboh) A, RbohB, RbohF and PR1 expression levels increased in inoculated resistant eggplant roots at an early stage (0–60 h postinoculation) and were at higher expression levels than those in susceptible eggplant roots. Ascorbate peroxidase, peroxidase and catalase activities were higher in inoculated resistant eggplant roots than in susceptible eggplant roots at the early stage. Hence, an early ROS burst positively regulates bacterial wilt resistance in eggplant.     

青枯病病菌对茄子相关防卫信号基因表达及活性氧含量的影响

本文以茄子抗青枯病自交系和感病自交系为试材,在接种青枯病菌后,对调控抗性反应的不同信号传导基因进行表达特性分析。结果表明,在9个基因中,EDS1、PAD4、NPR1、SGTI和WRYK70等5个基因的表达均随着接种诱导时间延长而增加,而且在抗病自交系‘E-31’中的表达量要高于感病自交系‘E．32’;而JAR1、NDR1、EIN2YeaRARl等基因的表达随着接种诱导时间的增加,表达水平变化不大。初步推断EDS1、PAD4、NPRI、SGTI和WRYK70可能与调节茄子抗青枯病抗性反应有关。同时茄子在接种后活性氧的含量均增加,但是感病植株的含量高于抗病植株。     

Role of hydrogen peroxide and ascorbate-glutathione pathway in host resistance to bacterial wilt of eggplant

Ralstonia solanacearum, a soil-borne bacterium causes bacterial wilt, is a lethal disease of eggplant (Solanum melongena L.). However, the first line of defense mechanism of R. solanacearum infection remains unclear. The present study focused on the role of induced H₂O₂, defense-related enzymes of ascorbate-glutathione pathway variations in resistant and susceptible cultivars of eggplant under biotic stress. Fifteen cultivars of eggplant were screened for bacterial wilt resistance, and the concentration of antioxidant enzymes were estimated upon infection with R. solanacearum. A quantitative real-time PCR was also carried out to study the expression of defense genes. The concentration of H₂O₂ in the pathogen inoculated seedlings was two folds higher at 12 h after pathogen inoculation compared to control. Antioxidant enzymes of ascorbate-glutathione pathway were rapidly increased in resistant cultivars followed by susceptible and highly susceptible cultivars upon pathogen inoculation. The enzyme activity of ascorbate-glutathione pathway correlates by amplification of their defense genes along with pathogenesis-related protein-1a (PR-1a). The expressions of defense genes increased 2.5?3.5 folds in resistant eggplant cultivars after pathogen inoculation. The biochemical and molecular markers provided an insight to understand the first line of defense responses in eggplant cultivars upon inoculation with the pathogen.     

芝麻营养生长期枯萎病抗性鉴定技术研究

芝麻枯萎病是芝麻主要真菌病害之一,由尖孢镰刀菌芝麻专化型(FOS,Fusarium oxysporum f.sp.sesami)引起,主要在苗期和成株期发生。为精准评价营养生长时期(2对真叶~现蕾)芝麻种质对FOS菌株的抗性水平,试验分析了FOS菌液浓度、蘸根接菌时间、致病力等条件下芝麻种质枯萎病病症及病情指数变化规律,建立了营养生长期芝麻枯萎病抗性精准鉴定方法。结果表明,1×10^6 cfu/mL^5×10^6 cfu/mL浓度下,植株蘸根接菌处理1~2周即可发病;第4周样本枯萎病病情指数趋于稳定。上述方法反映不同芝麻种质营养生长期对FOS菌株的抗性水平以及不同FOS菌株的致病力。营养生长期芝麻枯萎病发生可分为0~4级共5个等级。采用上述鉴定方法对42份芝麻种质进行抗枯萎病鉴定结果显示,野生种Sesamum radiatum Thonn.ex Hornem.高抗枯萎病(DI=0),而S.angustifolium(Oliv.)Engl.高感枯萎病(DI=100)。40份栽培种资源中,高感(HS)种质比例极高(55%),抗病种质比例较低(27.5%)。研究结果为深入开展芝麻抗枯萎病遗传机理分析提供了技术支持。     

Introgression of bacterial wilt resistance from eggplant to potato via protoplast fusion and genome components of the hybrids

Key message

Bacterial wilt resistant somatic hybrids were obtained via protoplast fusion between potato and eggplant and three types of nuclear genomes were identified in the hybrids through GISH and SSR analysis.

Abstract

Cultivated potato (Solanum tuberosum L.) lacks resistance to bacterial wilt caused by Ralstonia solanacearum. Interspecific symmetric protoplast fusion was conducted to transfer bacterial wilt resistance from eggplant (S. melongena, 2n = 2x = 24) into dihaploid potato (2n = 2x = 24). In total, 34 somatic hybrids were obtained, and of these, 11 rooted and were tested for genome components and resistance to race 1 of R. solanacearum. The hybrids exhibited multiple ploidy levels and contained the dominant nuclear genome from the potato parent. Three types of nuclear genomes were identified in the hybrids through genomic in situ hybridization (GISH) and simple sequence repeat (SSR) analysis, including (1) the potato type of the tetraploids in which eggplant chromosomes could not be detected by GISH but their nuclear DNA was confirmed by SSR, (2) the biased type of the hexaploids in which the chromosome dosage was 2 potato:1 eggplant, and (3) the chromosome translocation type of the mixoploids and aneuploids that was characterized by various rates of translocations of nonhomologous chromosomes. Cytoplasmic genome analysis revealed that mitochondrial DNA of both parents coexisted and/or recombined in most of the hybrids. However, only potato chloroplast DNA was retained in the hybrids speculating a compatibility between cpDNA and nuclear genome of the cell. The pathogen inoculation assay suggested a successful transfer of bacterial wilt resistance from eggplant to the hybrids that provides potential resistance for potato breeding against bacterial wilt. The genome components characterized in present research may explain partially the inheritance behavior of the hybrids which is informative for potato improvement.     

Transfer of fungal tolerance through interspecific somatic hybridisation between Solanum melongena and S. torvum

Tolerance to Verticillium wilt is found in a wild relative of eggplant, Solanum torvum. To transfer this tolerance to eggplant (S. melongena), protoplast fusions between eggplant and irradiated S. torvum protoplasts were performed. Putative hybrids were regenerated and evaluated for Verticillium tolerance and field value. A total of 12 plants were eventually selected to be incorporated into the breeding program, as they showed a high degree of Verticillium tolerance under field conditions combined with the morphological characters of eggplant, including normal seed set. DNA analysis showed one of these tolerant plants to have a changed pattern, with some changes corresponding to specific pattern characteristics of S. torvum. Received: 12 April 1998 / Revision received: 9 September 1998 / Accepted: 29 November 1998