水稻新品种(系)抗稻瘟病性鉴定研究

1986-1991年对17个省、市、自治区36个科研大专院校提供的804份水稻新品种(系)进行了水稻抗稻瘟病苗期和成株期鉴定,从中提出一批可供育种和生产上直接或间接利用的抗性材料。     

持续低温对水稻稻瘟病抗性的影响

研究了在持续低温作用下,具有不同抗性的品种汕优多系１号和汕优６３对稻瘟病菌的抗性变化。结果表明：（１）持续低温处理比未经处理的对照稻瘟病病斑要早出现２－３ｄ;低温处理３－５ｄ出现病斑数目最多,对病害发生最为有利;但处理时间过长,则不利于病害;（２）持续低温处理３－５ｄ对病斑扩展的影响显著。     

水稻外引品种（系）抗稻瘟病性鉴定

１９９１～１９９５年共鉴定由国际水稻所提供的国际稻瘟病圃材料１２３５份。通过苗期人工接种和田间自然诱发鉴定，筛选出ＩＲ１３７６１－１１－３、ＩＲ１８３４９－２２－１－２－１－１、ＩＲ３８７、ＧＩＺＡ１７５、ＭＩＬＹＡＮＧ９５、水源３５５６份抗性较为稳定的材料，可供育种和生产部门选用     

内生菌醇提取物对水稻抗稻瘟病的诱导作用

本文研究内生菌醇提取物WS对水稻稻瘟病的诱导抗性,在水稻3叶1心期喷施3种浓度WS,通过诱导抗病效果筛选WS最佳使用浓度,同时研究最佳浓度对水稻相关抗病防御酶活性的影响.结果表明,浓度50 ng/mL WS喷雾处理时诱导抗病效果最佳,病情指数降低12.89,诱导效果达到55.5％.能够显著提高PAL、PPO、几丁质酶活性,PAL活性高于CK处理30.28％,PPO、几丁质酶活性分别是CK处理的1.18、1.70倍.     

乙烯信号参与调控水稻纹枯病抗性的研究

 乙烯(ET)信号在植物抵抗各种逆境反应中具有重要作用,但目前关于乙烯信号在调控水稻纹枯病抗性中的作用仍缺乏系统研究。本研究以感纹枯病水稻品种Lemont和抗纹枯病水稻品种YSBR1为材料,分析了纹枯病菌接种后乙烯合成关键基因及信号传导途径中标志基因的表达水平,结果显示,纹枯病菌侵染可显著诱导相关基因的表达、激活乙烯信号。进一步采用乙烯合成抑制剂和信号激活剂处理水稻并进行接种鉴定,结果显示,两种化学剂分别可抑制或激活乙烯信号;无论是Lemont还是YSBR1,激活其乙烯信号均可显著增强抗病性,而抑制该信号均显著降低抗病性。对3个乙烯受体突变体(ethylene response2, etr2; ethylene response3, etr3; ethylene response sensor2, ers2)进行温室接种鉴定,发现突变体的病斑长度均极显著高于野生型对照。以上研究表明乙烯信号在水稻对纹枯病菌的防卫反应或基础抗性中具有十分重要的作用,结果将为进一步解析“水稻-纹枯病菌”间的互作机制、制定合适的病害综合防控策略提供理论基础。     

水稻空间诱变育种抗稻瘟病研究

粤香占和青华占2个水稻品种的种子经过高空气球搭载后,选育出株型好、分蘖力强,产量提高的空间诱变高代(6代以上)品系.为了解空间诱变品系对稻瘟病的抗性变异情况,采用36个广东省稻瘟病菌不同致病型的代表菌株分别对粤香占和青华占及其空间诱变品系进行抗谱测定.研究结果表明:①空间诱变能使水稻品种产生稻瘟病抗性变异,有的诱变品系比原种抗病,有的则比原种感病.②空间诱变产生的抗病性变异比较复杂,不同品种的抗病性变异不一样.③空间诱变引起的稻瘟病抗性变异有可能是由多个抗病基因引起的.④空间诱变可以获得抗稻瘟病的特异种质.粤香占是感病品种,本研究测定的抗谱为25%,从其空间诱变品系中,获得高抗稻瘟病的2个品系YX-8和YX-9,抗谱达到94.4%.空间诱变抗病品系的穗颈瘟鉴定试验正在进行中.     

对38个水稻新品种抗瘟性的评价

1986—1993年，对吉林省38个新选育及外引水稻品种，采用分菌系人工接种与病区多点异地生态病圃自然诱发方法，连续3—4年分别作了抗瘟性鉴定。据此拟定出《综合数年多点（菌）次鉴定结果评价水稻抗瘟性标准》（SDJPK），分别将参鉴新品种对苗、叶、穗瘟抗性作了评价。     

水稻品种抗病性变化与稻瘟病菌致病性变异的相关效应研究

 根据1979~1992年对四川省稻瘟病菌致病性与水稻品种抗病性的监测结果表明:稻瘟病菌种群变异与水稻品种抗病性变化密切相关。潜在致病小种随水稻种植面积比率的扩大而增殖,最后成为优势小种。     

四川省水稻抗稻瘟病育种现状及发展方向

稻瘟病是四川省水稻的主要病害，实践证明，选育种植抗病品种是控制该病害的最经济有效的措施。1981年开始，植物病理工作者与遗传育种工作者经过20年的协作攻关，研究制定了一套适合四川生态特点的有关病菌生理小种、水稻品种抗瘟性鉴定的规范化技术和评价体系，探明稻瘟病菌致病性变异和水稻品种抗瘟性丧失规律，筛选出200多份稻瘟病抗源材料，育成穗颈瘟1－5级的抗病品种（组合）27个，累计推广面积达2750万hm^2，有效地控制了稻瘟病的流行。在此基础上，提出今后重点研究领域和抗病育种策略。     

离体水稻叶片划伤接种鉴定稻瘟菌的致病型

抗瘟品种的培育和抗瘟基因布局需要快速、准确、大规模地定性水稻抗源及其后代的抗瘟基因型和稻瘟菌致病型。为此, 本研究建立了水稻离体叶片划伤接种方法。该方法依据主效抗瘟基因抵抗稻瘟菌在寄主体内扩展的特点, 通过针刺在水稻叶片上造成伤口, 避免了寄主侵入抗性的干扰, 从而有利于抗扩展性的定性鉴定。作者利用活体喷雾接种、叶片无划伤接种和本研究建立的离体叶片划伤接种等3种接种方法, 在秧苗4~6叶龄期, 对菌株12-DG-68在24个水稻抗瘟单基因系上的致病反应进行了测定, 结果显示：叶片划伤接种的检测结果稳定、一致; 而叶片无划伤接种和活体喷雾接种的检测结果假抗性比例分别为12.5%和4.2%, 不同叶龄期的叶片间反应型不一致率达7%。此外, 离体叶片划伤接种还可利用菌丝块接种, 以鉴定分生孢子产量低的菌株的致病型。因此, 水稻叶片划伤接种是一种准确、稳定和方便的稻瘟菌接种方法, 可用于大规模定性测定水稻抗源及其后代的抗瘟基因型和稻瘟菌的致病型。     

Assessment of blast disease resistance in transgenic PRms rice using a gfp-expressing Magnaporthe oryzae strain

Rice blast caused by the fungus Magnaporthe oryzae (anamorph Pyricularia grisea ) is one of the most devastating diseases of cultivated rice worldwide. In this study, a green fluorescent protein ( gfp )-expressing M. oryzae strain was generated and used to investigate the infection process in a commercial rice cultivar. Expression of the gfp gene did not affect the pathogenicity of the M. oryzae transformants. Confocal microscopy allowed in vivo imaging of this pathogen during infection of rice tissues. Magnaporthe oryzae pathogenicity was examined on both leaf and root tissues. In roots of wild-type plants, the fungus penetrated into epidermal and cortical cells, and colonized the central cylinder and xylem vessels. However, the dimorphic growth pattern typically observed during the biotrophic and necrotrophic stages of leaf colonization was not observed during colonization of root tissues. Furthermore, events occurring during infection of rice plants constitutively expressing the maize pathogenesis-related PRms gene were characterized and compared with those occurring during the interaction of this pathogen with untransformed rice plants. Fungal penetration was drastically reduced and delayed in tissues of PRms plants compared to untransformed plants. These results indicated that the gfp -expressing M. oryzae represents a strategic tool for the assessment of blast disease resistance in transgenic rice which can be also applied to the analysis of the M. oryzae interaction with other cultivars or mutants of important crop species.     

甘蓝夜蛾CYP9A90基因的克隆及溴氰菊酯对其诱导表达

P450 CYP9A家族基因与昆虫对拟除虫菊酯类杀虫剂的抗性相关。为证实溴氰菊酯对甘蓝夜蛾CYP9A基因的诱导效果,本研究采用RT-PCR和RACE技术克隆获得甘蓝夜蛾CYP9A基因,real-time PCR检测该基因在甘蓝夜蛾不同组织中的表达差异及溴氰菊酯处理甘蓝夜蛾5龄幼虫不同时间后该基因相对表达量变化,研究结果可为甘蓝夜蛾对溴氰菊酯的抗性治理提供依据。结果表明:克隆得到甘蓝夜蛾CYP9A基因cDNA全长序列,该序列包含1 828bp,包括1个125bp的5′非编码区,1个104bp的3′非编码区和1个1 599bp的开放阅读框,编码532个氨基酸,分子量约为61.1kDa,等电点为8.84,GenBank登录号为KR676343,被国际P450命名委员会命名为CYP9A90。Real-time PCR分析结果表明,该基因在甘蓝夜蛾5龄幼虫6个组织中表达情况不同,其中在脂肪体中表达量最高。低剂量溴氰菊酯作用后不同时间点CYP9A90mRNA总体呈现诱导表达趋势。     

Investigation of rice blast development in susceptible and resistant rice cultivars using a gfp‐expressing Magnaporthe oryzae isolate

In this study, an isolate of Magnaporthe oryzae expressing the green fluorescent protein gene (gfp) was used to monitor early events in the interaction of M. oryzae with resistant rice cultivars harbouring a blast resistance (R) gene. In the resistant cultivars Saber and TeQing (Pib gene), M. oryzae spores germinated normally on the leaf surface but produced morphologically abnormal germ tubes. Germling growth and development were markedly and adversely affected in leaves of these resistant cultivars. Penetration of host cells was never seen, supporting the idea that disruption of germling development on the leaf surface might be one of the resistance mechanisms associated with Pib function. Thus, this particular R gene appeared to function in the absence of host penetration by the fungal pathogen. Confocal laser scanning microscopy of M. oryzae‐infected susceptible rice cultivars showed the dimorphic growth pattern that is typically observed during the biotrophic and necrotrophic stages of leaf colonization in susceptible cultivars. The suitability of the gfp‐expressing M. oryzae isolate for further research on R‐gene function and identification of resistant genotypes in rice germplasm collections is discussed.     

浙江省水稻新品种(系)对稻瘟病的抗性研究

用4个稻瘟病菌小种对浙江省280份水稻新品种(系)进行稻瘟病抗性分析,结果表明:浙江省水稻新品种(系)对稻瘟病菌具有较好的抗性,籼稻和粳稻对参试菌株的全抗率分别为21.62%和39.05%,说明粳稻具有较籼稻更广的抗谱;浙江省新品种(系)对ZB13小种具有较高的感病率,籼稻和粳稻的感病率分别为69.37%和52.07%。     

Impact of tricyclazole and azoxystrobin on growth,sporulation and secondary infection of the rice blast fungus,Magnaporthe oryzae

BACKGROUND: Rice blast, caused by Magnaporthe oryzae B. Couch sp. nov., is one of the most destructive rice diseases worldwide, causing substantial yield losses every year. In Italy, its management is based mainly on the use of two fungicides, azoxystrobin and tricyclazole, that restrain the disease progress. The aim of this study was to investigate and compare the inhibitory effects of the two fungicides on the growth, sporulation and secondary infection of M. oryzae. RESULTS: Magnaporthe oryzae mycelium growth was inhibited at low concentrations of azoxystrobin and relatively high concentrations of tricyclazole, while sporulation was more sensitive to both fungicides and was affected at similarly low doses. Furthermore, infection efficiency of conidia obtained from mycelia exposed to tricyclazole was affected to a higher extent than for conidia produced on azoxystrobin‐amended media, even though germination of such conidia was reduced after azoxystrobin treatment. CONCLUSIONS: This study presents for the first time detailed azoxystrobin and tricyclazole growth–response curves for M. oryzae mycelium growth and sporulation. Furthermore, high efficacy of tricyclazole towards inhibition of sporulation and secondary infection indicates an additional possible mode of action of this fungicide that is different from inhibition of melanin biosynthesis. Copyright © 2012 Society of Chemical Industry     

水稻主要抗瘟基因对福建稻瘟菌群体的抗性分析

 用1995-2003年间在福建省水稻产区采集的稻瘟菌代表菌系的108个分离菌,它们在CO39近等基因系上测定被划分为30个毒性类型,用它们在30个水稻抗稻瘟病近等基因系或单基因系品种上进行抗病性测定。结果表明水稻抗稻瘟病基因Pi-k^h抗性最强,抗性频率高达98.15%,Pi-1和Pi-9(t)也具有较高的抗性频率,是较好的抗源;对2个和3个Pi基因的联合抗性频率的分析,发现一些联合抗性频率极高,甚至有达到100%的组合,表明抗瘟育种采用多个Pi基因聚合,易于获得抗性强的品种。根据抗病基因与供试菌株互作的亲和性,对供试30个Pi基因可能的系统关系分析得到的初步信息可为抗病基因的聚合与布局策略提供参考。     

251份水稻品种(系)对稻瘟病的抗性鉴定及抗性多样性分析

 采用稻瘟病菌22个鉴别菌株对251份水稻品种（系）进行了抗病性测定,并进行了聚类分析和基因型推导。抗病性测定结果表明,251份品种（系）对鉴别菌株表现出的抗谱类型存在明显的多样性。根据供试品种（系）对鉴别菌株的抗感反应型,可将其划分为12个组,各组的品种（系）对鉴别菌株的反应型和抗性频率存在明显差异。基因型推导结果表明,在251份品种（系）中,70份品种涉及14个抗瘟基因,181份品种（系）不含有待测基因中的任何一个,但都含有其他抗瘟基因。聚类分析结果与基因型推导结果不存在简单的一一对应关系。珍科等10份品种（系）可作为田间抗瘟性筛选的候选材料。