水稻品种抗黑条矮缩病人工接种鉴定方法

为建立科学有效的水稻品种抗黑条矮缩病人工接种鉴定方法,分别研究了病毒在介体体内的循回时间、接种时间、接种强度、水稻接种苗龄4个因素对鉴定效果的影响。结果显示,病毒在介体体内的循回时间为12~15天或21~24天条件下,鉴定效果优于8~11天和16~17天处理;接种48~72 h条件下,鉴定效果优于12~24 h处理;有效接种强度4~20虫/苗条件下,鉴定效果优于1~3虫/苗处理;水稻接种苗龄0.5~2.5叶龄条件下,鉴定效果优于2.5~3.5叶龄处理。由此构建了水稻抗黑条矮缩病人工接种鉴定方法:循回时间为12~15天、接种时间48~72 h、有效接种强度4~20虫/苗和水稻接种苗龄0.5~2.5叶。在此条件下对不同抗性表现的水稻品种进行鉴定,其鉴定结果与重病区田间鉴定没有显著性差异,表明所建立的人工接种鉴定方法能客观地反映水稻品种对黑条矮缩病的抗性水平。     

水稻抗纹枯病苗期快速鉴定技术研究

 以抗感反应不同的5个水稻品种为试验材料,在人工气候箱、控温室中进行水稻苗期抗纹枯病接种试验,并与田间相应的成株期抗性试验进行比较,研究水稻苗期快速鉴定技术。结果表明:85%的相对湿度为纹枯病菌侵染危害水稻苗期植株的适宜湿度;苗期5个水稻品种对纹枯病抗性差异极显著,可将其分为相对感病(Lemont、武育粳3号)和相对抗病(YSBR1、Jasmine85、特青)2大类;接种叶龄对发病程度有显著的影响,5个品种在四叶期接种时的平均病级显著高于五叶期接种的平均病级;苗期水稻品种间抗感差异小于田间鉴定试验结果,但两者间品种抗感趋势基本一致。苗期快速鉴定技术可用于大规模水稻品种(组合)的抗性筛选或初步鉴定。     

水稻稻曲病室内人工接种技术

为提高稻曲病人工接种的发病效果和稳定性,在温棚条件下采用水稻孕穗期注射接种法分别研究了稻曲病菌不同接种体、培养时间、接种浓度和接种时期的接种效果。采用病菌马铃薯蔗糖液体培养基（potato sucrose broth,PSB）作为接种体,其穗发病率为100%,明显好于病菌米糠培养液（23.33%）。病菌在PSB中培养5～7天接种效果较好,随病菌培养天数的延长,接种效果明显下降。接种的分生孢子浓度越低水稻病穗率和病粒数也越低。在水稻品种两优培九孕穗中后期,采用含分生孢子浓度为4×106个/mL的病菌PSB培养液注射接种,穗发病率达100%,平均病粒数35.1粒,最高达87粒。研究表明,温棚条件下建立的稻曲病人工接种技术能获得稳定的发病效果,并可区别水稻品种间的抗性差异。     

高效、准确的玉米粗缩病人工接种鉴定技术

为建立高效、准确的玉米粗缩病人工接种鉴定技术,通过将玉米人工接种水稻黑条矮缩病毒(Rice black-streaked dwarf virus,RBSDV),研究了温度对RBSDV在灰飞虱体内循回期的影响以及玉米接种苗龄、接种强度及接种时间对人工接种鉴定效果的影响。结果表明:病毒在灰飞虱体内的循回期随温度升高而缩短,在平均温度分别为18.50、23.50、27.50℃时,循回期分别为36.27、21.76、16.38 d;在玉米芽鞘期至2叶1心期接种,接种强度每苗为7~10头虫,接种时间72~96 h条件下,鉴定效果优于其它处理;感病对照郑单958、掖107、掖478发病充分,病情指数为97.70%~100.00%。用已知抗性水平的玉米自交系和杂交种进行验证,鉴定结果表明该人工接种鉴定技术可客观反映供试玉米材料的实际抗性水平。     

江苏省主栽水稻品种抗赤霉病鉴定及毒素积累分析

为明确江苏省主栽水稻品种赤霉病抗性水平,2021—2022年采用人工接种结合田间自然发病情况对16个水稻品种进行赤霉病抗性鉴定,并分析了水稻籽粒中单端孢霉烯族化合物积累水平。结果表明：在人工接种条件下,9个水稻品种(4个粳稻和5个籼稻)连续两年对赤霉病达到中抗水平,籼稻品种对赤霉病抗性水平显著高于粳稻品种(P<0.05);在自然发病条件下,不同品种对赤霉病抗性水平不仅在年度间存在明显差异,也与人工接种鉴定结果存在较大差异,籼稻品种对赤霉病的抗性同样高于粳稻品种。籽粒中单端孢霉烯族化合物积累水平与病情指数呈极显著正相关(P<0.01)。本研究揭示了江苏省部分主栽水稻品种赤霉病抗性水平,为水稻赤霉病抗性鉴定评价、抗赤霉病水稻品种的培育和利用提供了科学依据。     

剑麻抗斑马纹病鉴定技术研究

为了促进形成一套剑麻抗斑马纹病的鉴定技术,本研究对剑麻斑马纹病最适宜接种条件进行了探讨,并利用活体叶片接种和离体叶片接种两种方式,以抗性不同的8个剑麻种质为试验材料,在人工气候箱和恒温室中进行剑麻抗斑马纹病接种试验,并与大田抗病性进行比较.结果表明:针刺法接种,25～30℃保湿培养是该病最适宜的发病条件;两种接种方法都能比较准确地鉴定剑麻种质对斑马纹病的抗病性,而活体叶片接种较离体叶片接种鉴定抗病性更加准确.     

甘蓝根肿病的人工接种体系与抗源材料筛选

试验分别比较了不同接种方法、不同接种浓度以及不同土壤pH条件下人工接种甘蓝根肿病后甘蓝的发病情况,筛选出一套甘蓝根肿病苗期抗性鉴定与评价的最佳接种体系:最佳接种方法为菌土法;最佳孢子接种浓度为1.6×108个/g基质,最佳土壤pH为6.0。利用最佳人工接种体系,对东北农业大学甘蓝课题组提供的20份甘蓝材料进行抗性鉴定,其中抗病(R)材料3份,中抗(MR)材料9份,感病(S)材料7份,高度感病(HS)材料1份。     

涂抹接种法在水稻稻瘟病穗颈瘟鉴定中的应用研究

1990～1992年研究了涂抹接种法应用于水稻稻瘟病穗瘟鉴定的接种效果,穗瘟涂抹接种是采用画笔将混有10～20万/ml孢子液的2％羧甲基纤维素水溶液直接涂抹于穗颈节部。经与喷雾、注射和湿棉球包穗接种法比较,涂抹接种法具有侵染率高,接种用菌量少,症状明显,且接近自然发病条件等特点。用该方法接种98个浙江省水稻品种进行验证表明,涂抹接种法可以明确区分水稻品种穗瘟抗性的强弱。     

用无菌苗法鉴定油菜对菌核病的抗(耐)病性

快速、准确、有效的抗病性鉴定方法是抗病育种的基础和保障.目前,国内外鉴定油菜菌核病的抗(耐)病性的方法归纳起来主要有草酸鉴定法、人工气候室盆栽苗接种鉴定、大田成株期人工辅助接种鉴定和大田病圃或非病圃自然侵染鉴定法等.草酸鉴定法对鉴定材料的破坏性大,育种中应用仅供筛选抗草酸的单株或材料,且不能鉴定抗菌核病菌侵入的材料.人工气候室盆栽苗接种鉴定受空间或成本的限制,难以用于大量的筛选工作,大田成株期人工辅助接种鉴定和大田自然侵染鉴定既受季节的限制,也受环境因素的影响.因此,探寻更高效、准确的抗(耐)病性鉴定方法,仍是抗病育种的迫切需要.本文介绍了一种准确、快速、高效的鉴定新方法.     

稻曲病的接种技术研究

 探索在田间条件下进行稻曲病的人工接种技术。通过比较3种接种体发现,上年保存于-20℃的厚垣孢子不能引起发病;用PS液体培养获得的薄壁分生孢子可引起发病,孢子浓度越高引起病穗率越高;菌丝片段-分生孢子混合液接种发病最严重。病原菌的培养菌龄对接种效果有影响。在水稻的3个时期接种,以抽穗破口前6~9d的效果最好。在一天的3个时段接种,以下午4~6时的病情最重。在接种液里添加马铃薯煮汁可显著提高接种发病率。本文的接种技术能高效引发稻曲病,最高穗发病率100%,最大病情指数93.96,最多穗病粒数110粒。该接种技术可鉴别出品种抗病性的差异。     

水稻旱晚播“活体病株”接种法的研究及应用

作者针对水稻抗瘟育种中“旱地稻瘟病圃”易受气候生态等条件的影响,导致年际间发病的稳定性和能否发病的安全性,改进以往喷雾接种或利用病区自然诱发法,采用活体病株持续接种,和人为小气候控制等措施,完善成一套发病系数高且稳定,方法简便实用,病圃容量大,鉴定结果可靠性强的综合技术措施,在连续几年试验中鉴定筛选出一批利用价值较高的种质资源和抗病性较好的系统材料。     

Two‐component cultivar mixtures reduce rice blast epidemics in an upland agrosystem

The effect of two‐component rice cultivar mixtures on the control of rice blast disease was studied in three different experiments under rainfed upland conditions in the Madagascar Highlands. The mixtures involved a susceptible cultivar (either susceptible or very susceptible) and a resistant cultivar in different mixture arrangements (random or row mixtures) and with different proportions of the susceptible cultivar (50, 20 and 16·7%), which were compared to the susceptible cultivar grown in a pure stand. The effect of these mixtures on the incidence and severity of leaf and panicle blast was measured weekly, and on yield and yield components at harvest time. The mixture effect was more efficient in reducing disease with a proportion of 16·7% susceptible component than with a proportion of 50%. Blast epidemic was significantly reduced in all three experiments. However, under high blast pressure, there was no reduction in the disease by the end of the epidemic and yields of the susceptible cultivar were almost zero whatever the mixture. In two other experiments performed under lower blast pressure, disease incidence and severity were significantly lower in mixtures, and yields of the susceptible cultivars grown in mixtures were higher than those of their respective pure stands. Cultivar mixtures are a promising strategy that could contribute to a more sustainable cultivation of rice under upland conditions in the context of subsistence agriculture in Madagascar, where all cropping operations are manual.     

水稻品种抗瘟性诱导研究Ⅰ.非生物诱导物的筛选及诱导方法初探

 采用经反复筛选的化学药剂亚硒酸钠、放线菌酮、氯化钾、硫酸锰和中药材前胡、白芷的提取液浸泡水稻种子或进行叶面喷雾,可诱导稻株产生对稻瘟病的抗性。在人工喷雾接种条件下,对感病品种抗叶瘟的诱导效果为43%~93%;在注射接种条件下,抗穗瘟的诱导效果为29%~78%。供试药剂对感病品种和弱致病菌株的诱导效果优于对抗病品种和强致病菌株;药液叶面喷雾的诱导效果优于药液浸种。     

非亲和性稻瘟病菌对水稻的诱导抗性

为研究稻瘟病菌Magnaporthe oryzae不同菌株间的相互作用,选择与单抗性基因系水稻IRBL5-M （携带抗性基因Pi5）表现为亲和性的菌株HN52与非亲和性的菌株HN119为研究对象,将其单独或混合接种到单抗性基因系水稻IRBL5-M中,并通过荧光显微镜观察接种后水稻叶鞘的发病情况及病斑面积,测定接种后水稻内相关抗性基因OsWRKY45、OsNPR1、OsPR10、OsMAPK2的表达量以及活性氧的变化。结果显示,相较于单独接种亲和性菌株,混合接种后单抗性基因系水稻IRBL5-M病斑发病面积减少;混合接种中亲和性菌株HN52菌丝侵染能力降低,侵染菌丝细胞间扩展率显著降低73.13%;同时单抗性基因系水稻IRBL5-M中OsWRKY45、OsNPR1、OsPR10和OsMAPK2抗性基因表达量显著增加,水稻叶片中活性氧含量增加,表明在菌株混合侵染过程中,非亲和性菌株可通过激发水稻的抗性反应来降低亲和性菌株对水稻的侵染程度。     

Identification of blast resistance genes in 358 rice germplasms (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) using functional molecular markers

Rice blast, caused by the fungus Magnaporthe oryzae, is one of the most devastating diseases of rice (Oryza sativa L.), and neck blast is the most destructive phase of this disease. Although neck blast causes tremendous yield loss, little is known about the molecular mechanisms underlying the neck blast resistance. To address this issue, we collected 358 rice varieties from different ecotypes in China and assessed them for the neck blast resistance under natural conditions favoring disease development in Jining, Shandong Province. Our results showed that 124 (34.6%) and 234 (65.4%) varieties were resistant and susceptible to M. oryzae under natural field conditions, respectively. Among the 358 rice varieties that were screened for the presence of 13 major blast resistance (R) genes against M. oryzae by using functional markers, 259 varieties contained one to seven R genes. In addition, the relationship between the presence of R genes and the disease reactions was also investigated by integrative analysis of phenotyping and genotyping based on functional markers. Our results showed that the rice blast resistance gene Pi2 was significantly correlated with neck blast resistance. Furthermore, any of the 13 major blast R genes was absent from 32 rice varieties exhibited obvious neck blast resistance, which would be the potential materials for identifying novel neck blast R genes. Taken together, our findings provide insight into the distribution of the 13 major blast R genes in the tested Chinese rice germplasm resources, which will serve as a basis for developing rice blast resistant. Furthermore, 32 rice varieties exhibited neck blast resistance, but they did not harbor any of the 13 major blast R genes. In the future, these varieties may be used to identify novel neck blast R genes.     

稻瘟病为害损失研究

采用人工接种诱发和药剂控制相结合的方法,造成小区间稻叶瘟病和稻穗瘟的发病梯度系统监测病情,收获期结合病情程度进行单株考上区测产,并分别建立了依据稻叶瘟病,稻穗瘟病和二者综合的损失估计模型。就稻叶瘟病地水稻有效穗、穗粒数、千粒重和精米率的影响作了定量研究。另外,地稻穗瘟病在不同品种,不同侵染发病时间以及不同发病部位的为害影响进行了初探。结果表明,稻穗瘟病的为害程度在不同品种上存在一定差异,与侵染发病     

Molecular mapping of the new blast resistance genes Pi47 and Pi48 in the durably resistant local rice cultivar Xiangzi 3150

The indica rice cultivar Xiangzi 3150 (XZ3150) confers a high level of resistance to 95% of the isolates of Magnaporthe oryzae (the agent of rice blast disease) collected in Hunan Province, China. To identify the resistance (R) gene(s) controlling the high level of resistance in this cultivar, we developed 286 F(9) recombinant inbred lines (RILs) from a cross between XZ3150 and the highly susceptible cultivar CO39. Inoculation of the RILs and an F(2) population from a cross between the two cultivars with the avirulent isolate 193-1-1 in the growth chamber indicated the presence of two dominant R genes in XZ3150. A linkage map with 134 polymorphic simple sequence repeat and single feature polymorphism markers was constructed with the genotype data of the 286 RILs. Composite interval mapping (CIM) using the results of 193-1-1 inoculation showed that two major R genes, designated Pi47 and Pi48, were located between RM206 and RM224 on chromosome 11, and between RM5364 and RM7102 on chromosome 12, respectively. Interestingly, the CIM analysis of the four resistant components of the RILs to the field blast population revealed that Pi47 and Pi48 were also the major genetic factors responsible for the field resistance in XZ3150. The DNA markers linked to the new R genes identified in this study should be useful for further fine mapping, gene cloning, and marker-aided breeding of blast-resistant rice cultivars.     

Comparison between the resistance to blast in panicles exserted from the main culm and primary tillers as measured in six rice varieties

Three rice japonica varieties Jiajing3768, Xiushui09, and Xiushui63 and three rice indica varieties Qianyou930, Liangyoupeijiu, and Liangyou6326 were used to analyze the resistance to neck blast, comparing panicles excised from the main culm and from the first primary tiller. Disease incidence in the necks, disease incidence in the rachis nodes, lesion length in the necks, and number of conidia in the necks were measured after artificial inoculation of excised neck pieces. Resistance to neck blast was higher in the panicles excised from the main culm than from the first primary tiller for the varieties Jiajing3768, Xiushui63, and Liangyou6326. However, the opposite was true for Xiushui09 and Liangyoupeijiu. In Qianyou930, the type of panicles had no significant influence on the resistance to neck blast.     

28%三环唑·嘧菌酯悬浮剂对水稻稻瘟病的防治效果

为科学合理地选择和使用田间控制稻瘟病的杀菌剂,本研究通过室内苗期测定和大田试验,评价复配剂28%三环唑·嘧菌酯SC对水稻稻瘟病的防治效果。结果表明,28%三环唑·嘧菌酯SC对水稻苗瘟及穗瘟均有很好的预防和治疗效果。在温室人工控制条件下,施药后接种的防效高于接种后施药的防效,最高防效分别为97.35%和92.70%。在田间对穗瘟的最高防效达到91.11%,与对照药剂75%三环唑WP和250 g/L嘧菌酯SC的平均防效相当,可以在生产上推广使用,推荐有效成分用量为252～420 g/hm~2。