57份小麦微核心种质白粉病抗性评价

为筛选对小麦白粉病具良好抗性的种质资源,本研究采用人工接种感病的方法,评价了57份小麦微核心种质材料的苗期与成株期抗性。结果表明,育成品种复壮30和兴义4号在苗期与成株期均表现为免疫或近免疫,具有优良的全生育期抗性。农家品种酱麦与猪屎麦和育成品种会宁10号、甘麦8号与日喀则8号5个品种在苗期表现为高感或中感,成株期则表现为近免疫或高抗,具有典型的成株期抗性。     

81份春小麦种质资源抗条锈病评价

本研究结合苗期分小种接种和大田成株期自然诱发条件下对供试的81份春小麦资源进行抗条锈病鉴定,分析其抗条锈病情况、抗病类型及抗病性关系,结果表明:具全生育期抗条锈病类型的资源有7份,具成株期抗条锈病类型的资源有39份,有30份资源为含有成株期和对部分小种失去抗性的全生育期抗病性的抗性类型;苗期分别接种7个小种后,表现抗病的资源数为9~25份不等,表现中抗的资源为2~6份不等,表现感病的有50~69份不等,成株期表现抗病的73份,表现中抗的3份,感病的5份,即供试种质资源大部分表现为苗期感病,而成株期具有抗病性。聚类分析结果表明,该81份种质资源抗性聚类相对分散,遗传多样性水平较高。研究结果为小麦育种提供优良的抗条锈性资源。     

不同小麦品种资源苗期和成株期麦长管蚜抗性鉴定和分析

为明确小麦苗期和成株期与麦长管蚜抗性的关系,2010—2012年采用蚜量比值法对94个小麦品种进行了温室苗期人工接种和田间成株期自然感蚜麦长管蚜抗性鉴定。结果表明,43个品种连续3年在成株期对麦长管蚜表现不同程度的抗性,其中高抗品种2个,中抗品种8个,低抗品种33个;45个品种在苗期对麦长管蚜表现不同程度的抗性,其中高抗品种5个,中抗品种16个,低抗品种24个;30个品种在苗期和成株期同时表现抗蚜,占供试品种的31.91%,其中C273和兰麦（陕西柞水）在苗期和成株期均表现为高抗;小麦品种在温室苗期人工接种和田间成株期自然感蚜的蚜量比值呈极显著正相关,苗期和成株期麦长管蚜抗性表现一致的共有66个品种,占供试材料的70.21%。研究表明温室苗期人工接种是快速鉴定小麦品种麦长管蚜抗性的有效方法。     

61份大麦种质资源对蠕孢菌叶斑病的抗性鉴定

 麦根腐平脐蠕孢菌[Bipolaris sorokiniana(Sacc.)Shoemaker]引起的叶斑病是大麦上重要的病害叶部之一,在世界各大麦主要种植区均能造成严重的经济损失。本研究利用3个致病力不同的蠕孢叶斑病菌株Z 12010、Z 12014和Z 13010分别对来源于中国及国外引进的61份大麦种质资源进行苗期抗蠕孢菌叶斑病鉴定,田间接种Z 12014菌株进行成株期抗病性鉴定。在所鉴定的大麦种质中未发现在苗期或成株期对叶斑病免疫的材料。苗期抗叶斑病分析发现,品种之间对3个菌株的抗性存在明显差异,Varunda、Legacy、Tradition、09GW-01、09GW-07、08PJ-36、09PJ-39、垦啤麦7号、垦啤麦10号、驻大麦3号、S-4和中饲麦1号等12份材料对3个菌株表现为中抗至高抗反应型。7份材料成株期抗Z 12014菌株,其中Legacy、蒙啤麦3号、09PJ-39、垦啤麦9号、垦啤麦11号和龙啤麦3号等6份材料具有全生育期抗性特点,品种10-3苗期对菌株Z 12014感病,而成株期抗该菌株。     

甘薯品种对疮痂病的抗性及其机理分析

为查明我国南方甘薯主栽品种抗疮痂病的能力,用疮痂菌孢子悬液对16份品种作苗期接种与田间病情系统观察.结果表明,广薯79-15、潮薯1号等8份表现感病-中感;福薯26、广薯88-70等8份表现中抗-高抗;不同抗、感品种在田间表现长短不一的感病期.对6份品种藤蔓不同的部位和株龄的组织结构作显微测量与计数、接种试验以及糖含量测定显示,不同品种皮层组织结构差异决定对疮痂病的抗、感性;抗病品种藤蔓具有较厚的角质层,叶片气孔和幼嫩组织的腺鳞数目都较少,表现潜育期长,病斑少.且同一品种不同株龄皮层结构差异与感病程度间有密切关系,但其藤蔓糖含量与品种抗性之间不相关.     

黑龙江省水稻品种资源对水稻细菌性褐斑病的抗性鉴定

选育和应用抗病品种是防治水稻细菌性病害最经济有效的措施。本研究采用苗期喷雾法和针刺法对黑龙江省42个水稻主栽品种进行抗性鉴定。结果表明,利用针刺法鉴定的各水稻品种中表现中抗以上的品种有14个,其中高抗品种2个,对接种水稻品种病斑长度进行差异显著性分析,42个水稻主栽品种间抗性存在显著差异;利用喷雾法鉴定各品种,病级表现1级的品种有7个,占16.7%。对数据进行比较分析,针刺法和喷雾法对抗病品种鉴定的结果基本一致。本研究鉴定的2个高抗稻种资源对于抗细菌性褐斑病生产实践提供重要价值。因此,加大对水稻品种资源的深入研究,对实现水稻细菌性褐斑病的可持续控制有着重要意义。     

61份大麦种质资源对蠕孢菌叶斑病的抗性鉴定

 麦根腐平脐蠕孢菌[Bipolaris sorokiniana(Sacc.)Shoemaker]引起的叶斑病是大麦上重要的病害叶部之一,在世界各大麦主要种植区均能造成严重的经济损失。本研究利用3个致病力不同的蠕孢叶斑病菌株Z 12010、Z 12014和Z 13010分别对来源于中国及国外引进的61份大麦种质资源进行苗期抗蠕孢菌叶斑病鉴定,田间接种Z 12014菌株进行成株期抗病性鉴定。在所鉴定的大麦种质中未发现在苗期或成株期对叶斑病免疫的材料。苗期抗叶斑病分析发现,品种之间对3个菌株的抗性存在明显差异,Varunda、Legacy、Tradition、09GW-01、09GW-07、08PJ-36、09PJ-39、垦啤麦7号、垦啤麦10号、驻大麦3号、S-4和中饲麦1号等12份材料对3个菌株表现为中抗至高抗反应型。7份材料成株期抗Z 12014菌株,其中Legacy、蒙啤麦3号、09PJ-39、垦啤麦9号、垦啤麦11号和龙啤麦3号等6份材料具有全生育期抗性特点,品种10-3苗期对菌株Z 12014感病,而成株期抗该菌株。     

瓜类细菌性果斑病研究进展

瓜类细菌性果斑病是发生在甜瓜、西瓜等葫芦科植物上的一种严重的世界性病害,此病是典型的种传细菌性病害,病原为嗜酸菌属西瓜种(Acidovorax citrulli)。本文围绕瓜类细菌性果斑病菌的分离检测、致病机理、遗传多样性及防治等方面的研究进展作一概述,阐明了瓜类细菌性果斑病的研究现状。     

水稻品种对细菌性基腐病的抗性及病原细菌致病力分化的研究

用浸根接种法测定了国内外622个水稻品种对水稻细菌性基腐病(Erwinia chrysanthemiBurkholder et al.)的抗性。结果表明,高抗品种有127个,中抗品种210个,中感品种189个,高感品种96个。粳稻发病重于籼稻,国际水稻(IR系统)品种较为抗病,而斯里兰卡水稻(BG系统)品种较为感病。用来自四个省的10个水稻菌株和2个玉米菌株(E.chrysanthemi pv.zeae)在20个水稻品种上人工接种测定,资料经统计分析,结果发现不同地区的水稻菌株并无致病性分化现象;但不同水稻品种间的抗性差异显著,玉米菌株对水稻的致病力相对较弱。     

江西省细菌性条斑病菌的致病型划分和水稻抗性资源的鉴定

为探明江西省水稻细菌性条斑病菌(Xanthomonas oryzae pv.oryzicola,Xoc)的致病力分化水平和品种资源的抗病性,本研究利用IRBB5、IRBB14、IR24、IRBB4、IRBB21和金刚30等6个鉴别品种,将分离自江西6个稻区的129株Xoc菌株划分为C1~C9 9个致病型;发现仅C5对鉴别品种IRBB4、C7对鉴别品种IRBB21表现为强互作关系;不同稻区的Xoc优势种群呈现多样性。孕穗期抗性测定结果显示:124个籼稻品种中,对C2型菌株5-16表现为中抗的品种仅占8.06%,对C6型菌株08-3-2表现为抗的品种占14.5%;252个东乡野生稻单株中,1株单株(东113)对C2型菌株表现高抗,14个单株表现抗性。东乡野生稻苗期注射接种结果显示,苗期抗病性与孕穗期抗性呈显著相关性,但苗期抗病性普遍低于孕穗期抗性,仅东113表现为中抗。但东乡野生稻对于Xoc的抗病性明显强于栽培稻,从野生稻中寻找抗性资源将是培育抗病品种的理想途径之一。     

Characterization of Acidovorax citrulli strains isolated from solanaceous plants

Acidovorax citrulli is the causal agent of bacterial fruit blotch disease of cucurbits. Strains of this pathogen are distributed into two major groups: Group I strains have been mainly isolated from melon and other non-watermelon cucurbits, while Group II strains have been mainly recovered from watermelon. Here we report the characterization of strains T1 and EP isolated from diseased tomato and eggplant plants, respectively, and further confirmed to belong to A. citrulli species. Based on PCR, PFGE, and rep-PCR, these strains showed high similarity to the Group II strain 7a1. Sequencing and comparative analyses revealed that the genomes of T1 and EP aligned with that of the Group II model strain AAC00-1, over 97.88% and 99.22%, respectively. The virulence of T1, EP, and 7a1 determined on tomato, eggplant, and watermelon was similar and significantly higher than that of Group I strain M6. In contrast, M6 was more virulent on melon. Expression levels of seven virulence genes measured 24 hr after inoculation of tomato, eggplant, watermelon, and melon showed that the expression pattern was generally similar in strains 7a1, T1, and EP, whereas for M6 the expression was high only on melon. Overall, our results indicate that the solanaceous strains belong to Group II. To the best of our knowledge, this is the first study that reports characterization of A. citrulli strains isolated from solanaceous species. The fact that A. citrulli is able to naturally colonize and cause disease in non-cucurbit crops poses additional challenges for management of this important pathogen.     

兼抗麦长管蚜和大麦黄矮病毒的小麦种质田间鉴定筛选

为鉴定筛选兼抗麦长管蚜和大麦黄矮病毒(Barley yellow dwarf virus, BYDV)的小麦种质,采用自然感蚜/感病系数法,对36个外引和远缘杂交选育的小麦种质材料进行了2年的田间鉴定,并分析了感虫性与感病性的相关关系。结果表明,2年中均兼抗麦长管蚜和BYDV的种质仅有KOKIPPCAS、KOK、Amigo-3和PI137739共4个材料,占总鉴定材料的11.11%;对二者均敏感的有98-10-35q-9、186Tm39、Tam200e12-14a、Tam200(27)7、小偃22、西农1376和小偃6号共7个材料,占19.44%。其它材料仅抗虫或仅抗病,或仅在一年中表现抗病或抗虫,如材料98-10-30和98-10-35a8抗麦长管蚜,但对BYDV敏感;材料Tam200(13)G和PIG23(2)C感蚜,但对BYDV有抑制作用。BYDV发生普遍率(发病株率)和严重度(病情指数)与有蚜株率显著相关,严重度还与感蚜指数显著相关,但感病植株的病级均值与有蚜株率无显著相关性。表明自然界长期的进化和选择使许多抗病虫基因得以保存下来,但较多抗性基因只在抗病或抗虫的某一方面表现有效,需给予更多关注。     

Bacterial fruit blotch of melon: screens for disease tolerance and role of seed transmission in pathogenicity

Bacterial fruit blotch (BFB) of cucurbits, caused by Acidovorax avenae subsp. citrulli, is a serious threat to the watermelon and melon industries. To date, there are no commercial cultivars of cucurbit crops resistant to the disease. Here we assessed the level of tolerance to bacterial fruit blotch of various commercial cultivars as well as breeding and wild lines of melon, using seed-transmission assays and seedling-inoculation experiments. Selected cultivars were also tested in a greenhouse experiment with mature plants. All tested cultivars/lines were found to be susceptible to the pathogen, and most of them showed different responses (relative tolerance vs. susceptibility) in the different assays; however, some consistent trends were found: cv. ADIR339 was relatively tolerant in all tested assays, and cv. 6407 and wild lines BLB-B and EAD-B were relatively tolerant in seed-transmission assays. We also provide evidence supporting a strong correlation between the level of susceptibility of a cultivar/line and the ability of the pathogen to adhere to or penetrate the seed. To the best of our knowledge, this is the first attempt to assess melon cultivars/lines for bacterial fruit blotch response.     

瓠瓜果斑病症状及其病原菌的鉴定

为有效防控瓠瓜果斑病,自浙江省象山县田间采集具有典型果斑病症状的瓠瓜样本,对其进行病原菌分离、形态观察、致病性测定及分子生物学鉴定,并利用特异性引物PL1/PL2 PCR扩增和基质辅助激光解吸电离飞行时间质谱（matrix assisted laser desorption ionization time-of-flight massspectrometry,MALDI-TOF-MS）技术对其病原菌进行亚群鉴定。结果表明：瓠瓜果斑病田间典型症状是发病叶片和果实上病斑由水渍状小斑点逐渐发展为伴有黄色晕圈的褐色不规则病斑,果实腐烂、有臭味。通过菌体形态和培养特性观察、PCR鉴定、16S rDNA序列分析和7个看家基因的系统发育分析将其病原菌鉴定为西瓜噬酸菌Acidovorax citrulli。从瓠瓜上分离的菌株均属于西瓜噬酸菌亚群I,从西瓜上分离的菌株均属于西瓜噬酸菌亚群II。     

Genetic diversity analysis of Acidovorax citrulli in China

Acidovorax citrulli has become quite common in China. A collection of 118 strains of A. citrulli was made from throughout China and other countries to determine their genetic relatedness. Strains were identified as A. citrulli by pathogenicity, phenotypic characterization, and PCR. Genetic diversity was determined using pulsed field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). PFGE electrophoresis resulted in nine genotypes, which could be typed into two groups based on host; group 1 included strains mainly from melon and group 2 included strains mainly from watermelon. MLST analysis resulted in 73 sequence types (ST) among the 118 A. citrulli strains. All A. citrulli strains were typed into three groups: group 1 with 82 strains (including type strain Fc247), group 2 with 35 strains and group 3 a singleton (Fc380). Similar to PFGE results, group 1 included strains mainly from melon and group 2 included strains mainly from watermelon. The difference was the 10 watermelon strains (pslbtw1-3, 5–11) from Beijing grouped with melon strains of group 1 based on MLST, suggesting that these 10 watermelon strains had a close relationship with melon. Our study indicated that there was genetic differentiation among A. citrulli strains between watermelon and melon. Also, our study was the first attempt to compare PFGE and MLST on analyzing genetic diversity of A. citrulli strains and proved MLST could better distinguish A. citrulli strains.     

Pathways of bacterial invasion and watermelon seed infection by Acidovorax citrulli

This study explored the pathways of ingress of Acidovorax citrulli, the causal agent of bacterial fruit blotch of cucurbits, into watermelon seeds. Up until 7 days post‐inoculation (DPI), a significantly higher percentage of watermelon seeds was infected with A. citrulli when the bacteria were applied (c. 1 × 10⁶ colony‐forming units) to stigmas versus ovary pericarps of female flowers. Immunofluorescence microscopy showed that, with stigma inoculation, A. citrulli colonized style and ovary tissues by 1 DPI, and the bacteria co‐localized with pollen germ tubes in these tissues. With ovary pericarp inoculation, A. citrulli cells penetrated the epicarp and mesocarp tissues by 1 DPI but did not reach endocarp until 4 DPI. Finally, manual pollination followed by stigma inoculation led to >53% A. citrulli‐infected seed lots, while A. citrulli was not detected in seeds/ovules generated by stigma inoculation without pollination (chemically induced parthenocarpy). These results show that stigma inoculation results in faster colonization of watermelon ovules by A. citrulli than pericarp inoculation, even though there is no difference in the levels of infection in mature seeds. The data also indicate that pollen germ tubes play an important role in A. citrulli ingress into watermelon seeds via stigmas.     

几种生化物质与西瓜抗蚜性的相关性

为了初步探讨西瓜抗蚜性的营养与次生代谢物质防御效应,采用分光光度法测定并分析了几种生化物质与西瓜抗蚜性的相关性。结果表明,抗性品种黑皮、绿美人、黑美人和惠兰的游离氨基酸、脯氨酸和可溶性氮含量分别为2.22~2.27、0.23~0.26和1.88~3.03 mg/g,显著低于感性品种金美人、小玲、甜美人和花绿的3.24~3.83、0.67~0.87和5.26~5.82 mg/g;抗性品种的糖/氮比为5.38~7.94,显著高于感性品种;抗、感品种间可溶性糖和维生素C(Vc)含量无显著差异;瓜蚜为害4 d后的抗性品种叶组织中的总酚和单宁酸含量显著增加,感性品种显著降低;丙二醛含量增加幅度显著高于抗性品种。研究表明,西瓜品种抗蚜性与糖/氮比及总酚、单宁酸含量均呈显著正相关,与可溶性氮、游离氨基酸、脯氨酸及丙二醛含量均呈显著负相关,但与可溶性糖和Vc含量无显著相关性。     

High-throughput screening reveals small molecule modulators inhibitory to Acidovorax citrulli

Acidovorax citrulli is a seed-transmitted gram-negative bacterium that can cause substantial economic yield loss in watermelon and melon production worldwide. Four small-molecule libraries containing 4,952 compounds were selected for high-throughput screening against A. citrulli wild-type strain Xu3-14 by evaluation of growth inhibition. One hundred and twenty-seven molecules (2.5% hit rate) were identified as bactericidal or bacteriostatic against A. citrulli at 100 μM. Secondary screens indicated that 27 candidate compounds were more effective against A. citrulli Group II strains than Group I strains (classified using repetitive element PCR). Several compounds were inhibitory to other pathogenic bacteria, including Clavibacter michiganensis subsp. michiganensis and Xanthomonas campestris pv. campestris, but did not affect the growth of plant beneficial bacteria Pseudomonas fluorescens and Bacillus subtilis. More than half of the compounds did not inhibit germination of Arabidopsis or watermelon seeds. The effect of small molecules on A. citrulli seed-to-seedling transmission was evaluated by applying each compound to inoculated watermelon seeds and assessing seedling infection. Nine compounds were chosen for further investigation based on their reduction of percentage seedling infection and compiling scores on their specificity, sensitivity, and phytotoxicity obtained in the secondary screens. The five best compounds were selected (thiamphenicol, nadifloxacin, pipemidic acid, ciclopirox, and zinc pyrithione) for greenhouse tests and were found to effectively reduce the seed-to-seedling transmission of A. citrulli in both artificially and naturally infested seeds. These top five compounds provide a basis for future development of an A. citrulli-specific bactericide.     

西瓜噬酸菌不同菌株与甜瓜幼苗早期互作的转录组学分析

西瓜噬酸菌(Acidovorax citrulli)引起的瓜类细菌性果斑病(bacterial fruit blotch,BFB)是西瓜、甜瓜等葫芦科作物上毁灭性种传病害。本文利用RNA-seq技术,分析了2个西瓜噬酸菌菌株(pslb65和AAC00-1)分别诱导甜瓜感病品种IVF667幼苗6 h和12 h后的基因表达谱。结果表明,这2个菌株侵染寄主6 h后,分别检测到1 029和3 561个差异表达基因,其中上调基因分别为355和1 621个,下调基因分别为674和1 940个;与寄主互作12 h后,差异基因分别有2 397和1 875个,上调基因分别为903和898个,下调基因分别为1 494和977个。GO功能注释发现,pslb65与甜瓜幼苗互作的差异基因显著富集在生物学过程中的发育和代谢2个亚类,所占比例分别为32.92%和35.36%。在分子功能中转录因子所占比例较大,达到67.65%;AAC00-1与甜瓜幼苗互作后,差异基因显著富集在生物学过程中的转运和定位中,比例分别为23.84%和24.18%。在分子功能中氧化还原酶活性亚类所占比例高达17.59%。西瓜噬酸菌pslb65和AAC00-1侵染寄主6 h,Rboh、CaMCML、CDPK和FLS2等编码基因在植物-病原互作途径(csv04626)多为下调表达,植物-病原互作途径被抑制;12 h,pslb65-甜瓜互作途径相关基因多下调,AAC00-1-甜瓜互作途径相关基因多上调,推测此时植物-病原互作途径被AAC00-1激活,引起寄主的防御反应。西瓜噬酸菌pslb65和AAC00-1均可诱导植物发病,但在引起植物感病途径上略有不同,这可能与它们来自西瓜噬酸菌的不同亚群有关。最后,选取pslb65与寄主互作途径的6个基因进行qRT-PCR验证,结果与转录组测序结果基本一致。本研究初步揭示了西瓜噬酸菌2个不同菌株与寄主互作在转录水平上的表达差异,为研究甜瓜与不同菌株的互作机制差异奠定了基础。