感抗油菜近等基因系混播对根肿病发病率的影响

近年来,根肿病的迅速蔓延已对我国油菜生产造成严重威胁。课题组前期多年多点大田试验发现,感病、抗病品种混播可显著降低感病油菜品种根肿病发病率。为进一步探究感病、抗病品种播种间距对根肿病的防控效果,本试验采用穴盘栽培,设置感、抗根肿病油菜近等基因系不同播种间距(0、2、4、6 cm),出苗后7 d接种根肿菌,接种42 d后调查幼苗发病率、病情指数,并测定根系主要成分。结果表明:(1)与单播相比,感病、抗病品种同播时,抗病品种苗期发病率无明显变化,而感病品种苗期发病率显著下降,其发病率与感病、抗病品种播种间距密切相关,二者距离为0 cm时的发病率及病情指数均显著(P0.05)低于其他处理;(2)与单播相比,感病、抗病品种同播时,抗病品种和感病品种根中可溶性糖及可溶性蛋白质含量均显著降低,酸不溶木质素含量在感病品种中显著增加,而在抗病品种中呈现先增加后降低的趋势;(3)发病率和病情指数均与根系可溶性糖和可溶性蛋白质含量显著正相关(P0.01),发病率与可溶性糖和可溶性蛋白质含量的相关系数分别为0.797、0.403,病情指数与可溶性糖和可溶性蛋白质含量的相关系数分别为0.822、0.509。综上,油菜感、抗品种混播可以显著降低感病品种的根肿病发病率,且混播间距在0 cm时效果最好。该技术可以作为油菜根肿病防控的技术措施,在根肿病发病区域油菜生产上有着较好的应用前景。     

基于大白菜近等基因系的抗根肿病QTL QS_B3.1的验证

根肿病是影响大白菜生长发育及其产量的一个重要病害。本研究在前人定位出抗根肿病QTL QS_B3.1的基础上,以相同的抗根肿病芜菁自交系‘Siloga’为供体亲本,大白菜自交系‘91-12A’为受体亲本,通过对回交和自交群体的分子标记前景和背景选择,从BC4F3群体中筛选出遗传背景恢复为‘91-12A’,且只含有QS_B3.1位点的大白菜近等基因系。对BC4F3群体进行根肿病抗性鉴定,抗病植株与感病植株的分离比例为3:1,证明QS_B3.1抗病位点表现为显性单基因遗传。BC4F3群体的QS_B3.1连锁标记分析表明,抗病植株均含有QS_B3.1位点,不含QS_B3.1位点的植株均表现为感病,且QS_B3.1位点缩小在A3染色体的一个2.89 Mb区间内。这一发现为今后芸薹属作物的抗根肿病品种的选育以及该基因的精细定位和克隆提供了有力的依据。     

甘蓝型油菜抗根肿病KASP标记开发和利用

为了准确高效的筛选抗根肿病材料,提高甘蓝型油菜根肿病抗性育种效率。通过对甘蓝型油菜抗病亲本的Crr1基因与感病亲本中相应的同源基因LOC103834349进行测序,寻找SNP位点,针对第1 486-1 487上的非同义突变位点,开发了一套精准检测抗感根肿病基因型的竞争性等位基因特异性PCR(Kompetitive Allele Specific PCR,KASP)标记。利用该KASP标记对来自42个F_2群体的771个单株进行分型检测,其中315个单株含有纯合感病基因型GG,322个单株含有纯合抗病基因型AA,134个单株含有杂合等位基因型GA。其中对编号为2033的F_2群体中125株材料的KASP分型情况进行χ~2检测,其中纯合抗病基因型AA 30株,纯合感病基因型GG 33株,杂合基因型GA 62株,经χ~2检测,抗病材料与感病材料符合3∶1理论值,抗病纯合基因型、抗病杂合基因型与感病纯合基因型的比值符合1∶2∶1理论值,说明该抗病位点为显性单基因抗病位点。结合田间表型检测鉴定,AA分型和杂合的GA分型单株田间检测均为抗病表型,GG分型均为感病表型,KASP分型结果与田间鉴定结果一致。比对结果说明该KASP标记对根肿病抗、感植株进行正确分型,表明基于Crr1基因和LOC103834349的SNP位点开发的KASP标记可以准确高效的应用于油菜抗根肿病材料的分子标记辅助选择育种。     

油菜抗根肿病品种筛选试验研究

通过2009-2010年在云南根肿病重发病区保山市隆阳区板桥镇进行油菜抗根肿病品种筛选试验研究,对云南省近年来主栽的9个油菜品种和7个从国内各育种单位引进的品种进行根肿病抗性研究。结果表明：人工模拟苗期渍害明显加重根肿病为害,显著降低油菜最终收获产量。根肿病从苗期始为害油菜、苔期最重、成熟期为害发病率和病情指数均略有下降。供试的16个品种抗根肿病能力大体可分为3个类群,‘A35’、‘花油7号’和‘云油双1号’3个品种全生育期均表现出较强的抗根肿病能力,可作为云南根肿病常发区主导品种和育种材料使用。     

抗根肿病种质资源的筛选及油菜抗根肿病种质的合成

为了筛选十字花科抗根肿病种质,将抗性基因转育到甘蓝型油菜中,利用四川成都根肿菌4号生理小种对西北农林科技大学油菜课题组搜集提供的30份十字花科种质进行接种筛选,并对病情指数进行显著性差异分析、欧式聚类分析及发病率与病情指数的相关性分析。最终获得7份高抗(HR)材料,包括3份白菜、3份芜菁和1份甘蓝,2份中感(MS)材料,2份感病(S)材料和19份高感(HS)材料,其中高感材料均为油菜;这30份材料的平均发病率为69. 13%,平均发病指数为51. 45;抗、感材料病情指数之间存在极显著差异,欧式聚类分析与病情指数分析结果相一致,发病率与病情指数之间存在线性相关。将筛选获得的抗病芜菁德白瑞与感病甘蓝型油菜育种骨干亲本徐09进行远缘杂交,利用形态学、抗性鉴定、SSR分子标记等方法对远缘杂交的F1进行了真假杂种鉴定。结果发现,获得的3株F1杂种叶色浓绿,叶片肥厚,叶缘刺毛较少,叶型修长呈椭圆形,整体性状表现介于两亲本之间且偏向于母本,接菌后无发病症状,并用筛选得到的15对具有较好扩增多态性的SSR引物扫描3株杂种及双亲,带型均同时表现父母本带型,最终确定这3份F1杂种均为真杂种。筛选的7份高抗材料和3份真杂种材料可以为根肿病抗性机理研究及油菜抗根肿病新品种的选育提供材料基础。     

西南地区油菜品种根肿病抗性及布局分析

为明确西南地区主栽油菜品种根肿病抗性,以期为合理品种布局提供依据,降低根肿病的危害和损失。结合病圃根肿病菌生理小种鉴定,在四川安县、大邑、广汉三地区,采用病圃自然发病法,对80个油菜品种进行根肿病抗性评价,并对22个品种连续3年进行抗性跟踪。结果表明：三地区病圃根肿菌为存在基因型分化的4号生理小种;供试油菜品种中无免疫根肿病品种,安县、大邑、广汉三地感病品种分别占比88.75%、83.75%、87.50%,其中55个品种在三地区均表现感病,即不适宜在根肿病菌为4号生理小种区域种植;抗性跟踪评价显示不同品种的抗性稳定性存在差异。‘浙油50’、‘油罐罐’适宜安县地区种植,‘黄金荚’、‘金油858’、‘油研9号’、‘种都油998’和‘渝黄4号’适宜大邑地区种植,‘志远油8号’适宜广汉地区种植。‘绵丰油5号’、‘德名油1号’表现抗性丧失趋势;‘丰油精’、‘高油48’抗性不稳定;‘矮架早’抗性完全丧失。     

芝麻种质资源抗茎点枯病鉴定与评价

１９９２～１９９４年,通过人工接种,在田间对“七五”期间初筛出的抗茎点枯病材料３２份进行抗茎点枯病的进一步鉴定。结果表明,无免疫品种、高抗品种和高感品种占比例低,抗病和感病品种占比例大。鹫丹山－３、发芝麻和狼尾巴抗性比较稳定。迟芝麻抗性较好,并且产量较高     

抗根肿病甘蓝型油菜新品种华油杂62R的选育

我国油菜根肿病发病面积在66.7万公顷左右,约占总生产面积的10%,已严重威胁到油菜的安全生产。基于此,本研究以含有CRb抗根肿病位点的大白菜材料CR Shinki为供体亲本,以甘蓝型油菜国审杂交种华油杂62的父本Pol.CMS恢复系Bing409为受体亲本,通过杂交、回交及自交等育种程序,结合前景和遗传背景筛选,将CRb抗病位点导入到Bing409中。在BC3F2代获得了遗传背景高度接近Bing409且含CRb抗病位点的抗根肿病新恢复系Bing409R,进而成功选育了我国首个抗根肿病杂交油菜新品种华油杂62R。CRb抗病位点在甘蓝型油菜背景中表现为单基因显性遗传,根肿病抗性遗传改良的同时并未对Bing409R及由其配制的杂交种华油杂62R的产量、品质造成不良影响,Bing409R及华油杂62R对我国四川、湖北、安徽等地区根肿菌生理小种具有免疫抗性。本研究的开展为我国油菜抗根肿病育种提供了宝贵资源,为我国抵抗油菜根肿病的威胁提供了重要保障。     

成都平原主栽大白菜品种根肿病抗性的鉴定

为明确成都平原地区主栽大白菜品种对根肿病的抗性情况,于2018年分别在成都市新都区、彭州市和崇州市3个试验田块对45个大白菜主栽品种采用田间抗鉴圃自然感染诱发法进行根肿病抗性鉴定,通过调查根部根肿病发病程度评价各品种的抗性。结果表明,在崇州自然病圃中表现为免疫的品种有4个,占总品种的8.9%;其余品种均表现为感病或高感,占比91.1%。彭州的结果显示,在所测定的45个品种中,免疫品种有6个;耐病品种有11个;剩余28个品种为感病或高感品种。而在新都发病田块中,只有2个品种表现为对根肿病免疫,16个品种表现为耐病,27个品种表现为感病或高感。部分品种在不同地区抗性结果表现不一致,试验鉴定到2个对根肿病免疫的优异品种,可为选育和应用抗性品种提供借鉴。     

核盘菌诱导下甘蓝型油菜防御相关基因表达差异分析

菌核病是油菜主要病害, 至今尚未在油菜及相关植物中找到抗性基因。本研究利用qRT-PCR法比较了抗病品种宁RS-1和感病品种APL01在接种核盘菌后0~48 h内11个防御相关基因的表达差异, 以揭示抗病品种宁RS-1的抗病机制。结果表明, 4个基因(PGIP、Cu/ZnSOD、OXO和GLP)在核盘菌诱导前后抗、感品种内表达量均较高, 且抗病品种的表达量显著高于感病品种, 尤其是PGIP基因, 抗病品种宁RS-1在24 h的表达量为诱导前的170.4倍, 而感病品种仅为诱导前的3.5倍, 该时期抗病品种PGIP的表达量为感病品种的1299.4倍;2个基因(LOX2和PDF1.2)在诱导前后抗、感品种内的表达量均较低, 但抗、感品种间表达量差异显著;5个基因(FeSOD、PAL、EDS1、PR1和EIN3)诱导前后抗、感品种内的表达量均较低, 且抗、感品种间的表达量差异不显著。推测抗病品种宁RS-1对菌核病的抗性可能是由于PGIP的上调表达, 抑制了核盘菌PG蛋白对侵染部位油菜组织细胞壁的降解, 从而抑制了油菜菌核病的发生与蔓延。     

抗根肿病甘蓝型油菜新品种华油杂62R的选育

The rapeseed clubroot disease incidence in China is about 0.67 million hectare, accounting for 10% of the canola production area, which become a serious threat for the safety of Brassica napus industry. Based on this, we used CR Shinki, a Chinese cabbage material containing CRb clubroot disease resistance locus, as the donor parent, and Pol.CMS restorer line Bing409, the parent of Brassica napus national approved varieties Huayouza 62, as the recipient parent, and the CRb resistance locus was introduced into Bing409 by breeding programs such as crossing, backcrossing, self-cross with the foreground and genetic background selection. In the BC₃F₂ generation, a new restorer line Bing409R with a genetic background close to Bing409 containing CRb resistance locus was obtained, and Huayouza 62R, the first rapeseed hybrid resistant to clubroot disease in China was successfully developed. The results were as follows: CRb disease resistance locus appeared as a dominant single-gene inheritance in B. napus background, and the genetic improvement of resistance to clubroot disease did not at the expense of yield and quality losses for new restorer line Bing409R and its hybrid Huayouza 62R. Bing409R and Huayouza 62R were showed immune-resistance to physiological races of Plasmodiophora brassicae in Sichuan, Hubei, and Anhui provinces in China. This study will provide valuable resources for the breeding of rapeseed in China, and supplemented important support to overcome the threat of rapeseed clubroot disease.     

油菜苗期耐盐碱性综合评价与根际土壤有机酸含量比较

为筛选耐盐碱油菜种质,研究有机酸在油菜抗盐碱过程中的生理作用,利用碱性盐（摩尔比为2:1的NaHCO₃和Na₂CO₃的混合物）进行处理,用模糊数学隶属函数分析法对8个油菜种质进行耐盐碱性综合评价,探究碱性盐耐性最强和最弱的油菜种质的根际土壤有机酸含量。结果表明,耐盐碱性综合评价为华油杂62>华双5号>崇2>16-P22外>青杂9号>金油158>17崇1>16-P32外。在碱性盐胁迫下耐性最强种质华油杂62和最弱种质16-P32外的根际土壤中检测到草酸、酒石酸、甲酸、苹果酸、抗坏血酸、乙酸、柠檬酸及琥珀酸8种有机酸。在碱性盐胁迫下,油菜根际土壤中草酸的积累量最高,抗坏血酸的积累量很低。华油杂62的各处理根际土壤有机酸含量总体高于16-P32外。     

Development of novel clubroot resistant rapeseed lines (Brassica napus L.) effective against Japanese field isolates by marker assisted selection

Clubroot is an important disease infectible to cruciferous plants and a major threat to rapeseed production in Japan. However, no clubroot resistant rapeseed cultivars have been released. We surveyed pathotype variation of six isolates collected from rapeseed fields and found they were classified as pathotype groups 2 and 4 using Japanese F₁ Chinese cabbage cultivars. We produced the resynthesized clubroot resistant Brassica napus harboring two resistant loci, Crr1 and Crr2, by interspecific crossing and developed resistant rapeseed lines for southern and northern regions by marker-assisted selection and backcrossing. We improved the DNA marker for erucic acid content to remove linkage drag between Crr1 and high erucic acid content and successfully selected lines with clubroot resistance and zero erucic acid for northern regions. A novel line, ‘Tohoku No. 106’, suitable for southern regions showed stable resistance against all six isolates and high performance in infested fields. We conclude that Crr1 and Crr2 are important genes for CR rapeseed breeding and marker-assisted selection is effective in improving clubroot resistance.     

甘蓝型油菜品系一些酶的活性与抗菌核病的关系

以6个抗病品种（系）M004、湘油15号、中R-888、中R-783、084、085和二个感病品种（系）98C04、甘油5号为材料,研究接种菌核病菌前后叶片中过氧化物酶（PO）、多酚氧化酶（PPO）、苯丙氨酸解氨酶（PAL）、超氧物歧化酶（SOD）的活性变化。结果表明：接种前,PO、PAL二种酶活性与油菜品种抗病性相关不显著,而PPO、SOD酶活性     

Evaluation of French Brassica oleracea landraces for resistance to Plasmodiophora brassicae

A total of 240 kale, 38 cabbage and 126 winter cauliflower French landraces from the B. oleracea genepool of INRA were assessed for resistance to clubroot caused by Plasmodiophora brassicaeWoron. Two French isolates of the pathogen (K and SJ) were used in the experiments under controlled conditions. The reaction of the 126 cauliflower accessions to naturally occurring clubroot was also evaluated in field trials. Kales exhibited considerable variation for expression of disease resistance and high levels of resistance were found in several accessions. In this group, single resistant plants were observed in most of the morphological types and from quite different geographical origins. Cabbage accessions were moderately to highly susceptible to both isolates. All cauliflower populations proved to be highly susceptible to K isolate and moderately susceptible to SJ isolate. In field trials, cauliflowers were also severely infected. Two lines selected from a resistant kale population were highly resistant against a large range of pathotypes of the pathogen. These lines presented a sufficient level of resistance to be directly useful in the breeding program in order to develop cauliflower and broccoli hybrids resistant to clubroot. This revised version was published online in July 2006 with corrections to the Cover Date.     

甘蓝型油菜对华北坝上冷凉环境的适应性

针对华北坝上冷凉区短季类经济作物缺乏,芥菜型油菜品质差、产量低等问题,于2017-2018年在农业农村部张北农业资源与生态环境重点野外观测试验站,引种了甘蓝型双低油菜品种华油杂62,以当地传统芥菜型油菜品种大黄为对照,监测了两品种的叶面积指数、角果净光合速率日变化、地上部干物质积累量、养分积累量和油菜农艺性状、产量等。结果表明,华油杂62较大黄生育期长约10d,平均叶面积指数高23.36%~36.73%;角果净光合速率不及大黄,N、P、K积累量高于大黄;干物质快速增长期延长21d,干物质积累潜值高46.86%;菜子产量较大黄高22.8%~54.0%。甘蓝型油菜华油杂62对华北坝上冷凉区的地理气候环境有较好的生态适应性,具有替代区域传统芥菜型油菜的优势。