Resistance to powdery mildew (Blumeria graminis f.sp. avenae) in oat seedlings and adult plants

In this work, 165 Avena sativa and Avena byzantina accessions were screened for resistance to powdery mildew caused by Blumeria graminis f.sp. avenae and the defence mechanisms of resistant plants were further characterized. Ten resistant and moderately resistant accessions were selected according to macroscopic assessment. A detailed histological study of selected genotypes showed a range of defence mechanisms, acting alone or in combination, that impeded fungal development at different stages. Since the resistance observed in the collection was scarce, a study of adult plant resistance was carried out in 45 genotypes selected from field trials. Nine oat landraces and two commercial varieties showed very high levels of adult plant resistance. A detailed study of the components of the adult plant resistance revealed a high increase of penetration and post‐haustorial resistance in the fifth compared to the first leaves. Identification of the resistance sources and characterization of underlying defence mechanisms will be useful for future breeding programmes and for further cellular and molecular studies to unravel the genetic basis of resistance, in this species in particular and in cereal–powdery mildew interactions in general.     

冀西北地区燕麦主栽品种（系）抗秆锈病鉴定

2011-2012年,在2个试验点,采用孕穗期接种混合菌种的方法,对冀西北地区40个燕麦主栽品种（系）抗秆锈病进行了鉴定。结果表明：9个皮燕麦品种中,‘冀张燕5号’、‘冀张燕2号’、‘坝燕1号’、‘冀张燕1号’和‘定燕1号’等5个品种,31个裸燕麦品种（系）中,‘白燕2号’、‘白燕11号’、‘冀张莜6号’、‘冀张莜12号’、‘花晚6号’、‘晋燕14号’和‘燕科1号’等7个品种,至少在1个试验点表现免疫、抗病或慢病,其中大多在温度较低的试验点1更抗病,认为与大多数燕麦抗秆锈病基因表现低温抗病性有关;其余28个品种（系）均表现感病或高度感病。     

拟南芥的抗病信号传导途径

 拟南芥是研究植物与病原物相互作用的模式植物。植物感病和抗病取决于病原物无毒基因产物和寄主抗病基因产物的识别,以及随后的相关防卫反应的激活。在拟南芥的抗病过程中,水杨酸、茉莉酸、乙烯等信号分子都不同程度地参与着抗病过程中的不同环节,起着非常重要的作用。由于这些信号分子在对不同病原菌的抗性中的作用存在差异,因而将抗病信号传导分为依赖于水杨酸和依赖于茉莉酸/乙烯的途径。本文将着重讨论这些信号分子在植物系统获得抗性以及诱导系统抗性中的作用。     

The resistance to powdery mildew (Oidium lycopersicum) in Lycopersicon species is mainly associated with hypersensitive response

The cultivated tomato ( Lycopersicon esculentum) is susceptible to powdery mildew ( Oidium lycopersicum). Six accessions of three related Lycopersicon species show high levels of resistance (Lindhout et al., 1994b). The present research aimed at describing the development of O. lycopersicum on susceptible cv Moneymaker and characterizing the defence response to O. lycopersicum in Lycopersicon accessions by histological analysis. Spore germination and (primary) haustorium formation in resistant accessions were as frequent as in the susceptible L. esculentum cv Moneymaker. A high frequency of necrosis of epidermal cells in which a haustorium was formed appeared to be the major defence response, indicating that resistance to O. lycopersicum in the Lycopersicon genus was predominantly based on the hypersensitive reaction. However, the resistance in L. parviflorum was less associated with hypersensitivity than in other resistant accessions, suggesting the existence of a different but still unknown resistance mechanism. In addition, evidence is provided that the level of resistance could depend on the genetic background and the plant age.     

异源Harpin对软腐欧氏杆菌与植物互作关系的影响

 本文通过三亲交配将带有梨火疫欧文氏杆菌功能完整的hrp基因簇的重组粘粒pCPP430转移到了胡萝卜软腐欧氏杆菌(Erwinia carotovora) Se9R中。从接合子中提纯的质粒与pCPP430的大小、酶切图谱相同;以该质粒为模板,用针对hrpN序列的引物经PCR扩增得到与hrpN大小相同的片段;用地高辛标记的hrpN作为探针进行Southern blotting分析,在接合子中的质粒上得到预期的杂交带。接合子可以在烟草叶片上稳定地引起过敏反应。另外,接合子可以在寄主马铃薯叶片上引起过敏反应样枯斑,表明异源harpin的表达及功能不受软腐菌的抑制。该接合子在体外产生的果胶酸裂解酶活性与受体菌Se9R无明显差别。低浓度的接合子在马铃薯块茎上的软腐症状明显低于受体菌Se9R。外源harpin表达可能不改变软腐欧氏杆菌的致病力、而是通过诱导马铃薯抗性达到减轻发病的作用。     

Signal Transduction in Resistance to Plant Viruses

Salicylic acid is part of a signal transduction pathway that induces resistance to viruses, bacteria and fungi. In tobacco and Arabidopsis the defensive signal transduction pathway branches downstream of salicylic acid. One branch induces PR-1 proteins and resistance to bacteria and fungi, while the other triggers induction of resistance to RNA and DNA viruses. This virus-specific branch can be activated using antimycin A and cyanide, or inhibited with salicylhydroxamic acid, suggesting a role for alternative oxidase in resistance to viruses. The virus-specific defensive pathway activates multiple resistance mechanisms. In tobacco, salicylic acid induces resistance to systemic movement of cucumber mosaic virus but has no effect on its replication or cell-to-cell movement. However, in the case of tobacco mosaic virus in tobacco, salicylic acid appears to induce interference with the synthesis of viral RNA.     

Staying alive – is cell death dispensable for plant disease resistance during the hypersensitive response?

Probably the most known and best studied type of plant resistance to pathogenic infections is the hypersensitive response (HR), a form of localized programmed cell death associated with restriction or killing of pathogens that often leads to macroscopically visible localized tissue necrosis. It is generally assumed that cell death and resistance within the HR are physiologically and genetically linked. However, there has been considerable speculation about whether cell death is an absolute requirement for resistance conditioned by the HR. This review discusses the relation of cell death and resistance in the HR, in particular, the importance of cell death in this process. We intend to focus on the increasing amount of research evidence showing that in several plant-pathogen interactions, the two main components of the HR – resistance and cell death – can be physiologically, genetically and temporally uncoupled. In other words, HR should be considered as a combination of resistance and cell death responses, where cell death may be dispensable for plant disease resistance. The varying contribution of these two components (i.e. cell death and resistance) generates an array of defense strategies differing in efficiency. Thus, a very early and rapid defense response seems to contribute to the development of macroscopically symptomless (extreme) resistance, while a moderately early defense response results in resistance with the concomitant development of controlled and limited cell and tissue death (HR). Accordingly, a delayed and failed attempt by the host to elicit resistance responses would result in massively stressed plant tissues (e.g. “systemic HR”) and a partial or almost complete loss of control over pathogen invasion. The dynamic nature of resistance responses in plants implies that resistance can be effective with or without cell death but its outcome and efficiency may depend primarily on the timing and speed of the host response.     

Comparative microscopic and molecular analysis of Thatcher near‐isogenic lines with wheat leaf rust resistance genes Lr2a,Lr3, LrB or Lr9 upon challenge with different Puccinia triticina races

Thatcher near‐isogenic lines (NILs) of wheat carrying resistance gene Lr2a, Lr3, LrB or Lr9 were inoculated with Puccinia triticina races of virulence phenotype BBBD, MBDS, SBDG and FBDJ. Puccinia triticina infection structures were analysed under the fluorescence microscope over a course of 14 days after inoculation (dai). The relative proportion of P. triticina and wheat genomic DNA in infected leaves was estimated with a semiquantitative multiplex PCR analysis using P. triticina‐ and wheat‐specific primers. The occurrence of a hypersensitive response (HR), cellular lignification and callose deposition in inoculated plants was investigated microscopically. In interactions producing highly resistant infection type (IT) ‘0;’, a maximum of two haustorial mother cells per infection site were produced, and there was no increase in the proportion of P.  triticina genomic DNA in infected leaves, indicating the absence of P. triticina growth. In comparison, sizes of P. triticina colonies increased gradually in interactions producing moderately resistant IT ‘1’ and ‘2’, with the highest proportion of P. triticina genomic DNA found in leaves sampled at 14 dai. In interactions producing susceptible IT ‘3–4’, the highest proportion of P. triticina genomic DNA was found in leaves sampled at 10 dai (45·5–51·5%). HR and cellular lignification were induced in interactions producing IT ‘0;’ and ‘1’ at 1 dai but they were not observed in interactions producing IT ‘2’ until 2 dai. No HR or cellular lignification were induced in interactions producing susceptible IT ‘3–4’. Furthermore, a strong deposition of callose was induced in Lr9 + BBBD and Lr9 + FBDJ (IT ‘0;’), whereas this defence response was not induced in resistant or susceptible interactions involving Lr2a, Lr3 or LrB, indicating that Lr9 mediated resistance was different from that conditioned by Lr2a, Lr3 or LrB.     

Engineered Resistance Against Fungal Plant Pathogens

Development of genetic engineering technology and molecular characterization of plant defense responses have provided strategies for controlling plant diseases additional to those based on chemical control or classical breeding programs. Most of these alternative strategies are based on the overproduction of one component of the plant's own defense response. Some strategies exploit the hypersensitive response, a rapid, localized death of tissue surrounding the infection site, which is observed in many resistant plants upon unsuccessful pathogen attack. Most approaches to increase resistance to fungi have been described to be successful under laboratory conditions. Incorporation of these successful, alternative strategies in resistance breeding programs of agriculturally important crops will depend on the results obtained from field experiments.     

Temperature requirements for germination,germ tube growth and appressorium formation of urediospores of Hemileia vastatrix

Requirements for germination, germ tube growth and appressorium formation of urediospores ofHemileia vastatrix, the causal organism of coffee leaf rust, were investigated by applying treatments with constant temperatures, ranging from 10 to 31°C, and with variable temperatures. Observations were made after 4, 6, 8 and 24 h of incubation of the spores in distilled water on glass slides. The lower and upper limits for germination were estimated to be 13 and about 30°C, respectively. Germination was quickest at 22 to 28°C, whereas appressoria formed more rapidly at 13 to 16°C. After 24 h of incubation, a broad optimum, from 16 to 28°C, was observed for germinationn and total appressorium formation. The shape of the appressoria was torpedo-like or roundish at 13 to 19°C, whereas at higher temperatures their shape was predominantly irregular. Germ tube length increased linearly with time, with an optimum at 19 to 22°C. The degree of branching of the germ tube was positively correlated with germ tube length. Germination percentage and appressorium formation were higher on leaf disks than on glass slides. With variable temperatures, significant correlation were found between temperature sum and percentage of germination, appressorium formation and germ tube length.The results show that germination and appressorium formation can occur at lower temperatures than reported in literature. Penetration byH. vastatrix seems te be realized most rapidly in nature, if, after wetting of the spores, a decline in temperature occurs from initially ca. 23 to 17°C within a few hours. Such conditions prevail during evening and early night in many areas where Arabica coffee is grown.Samenvatting De temperatuurbehoeften voor kieming en appressoriumvorming van uredosporen vanHemileia vastatrix werden onderzocht door incubatie bij constante temperaturen, 10 tot 31°C, en variabele temperaturen. Waarnemingen werden gedaan na 4, 6, 8 en 24 uur incubatie van de sporen in gedestilleerd water op glasplaatjes. De onder- en bovengrens voor kieming bleek bij 13 en ca 30°C te liggen. De kieming kwam het snelst op gang bij 22 tot 28°C en de appressoriumvorming bij 13 tot 16°C. Na 24 uur incubatie werd een breed optimum waargenomen van 16 tot 28°C voor het kiempercentage en het totale aantal appressoria. De vorm van de appressoria was torpedoachtig of rond bij 13 tot 19°C, terwijl bij hogere temperaturen de vorm meestal onregelmatig was. De lengte van de kiembuis nam vrijwel rechtlijnig toe met de tijd, met een optimum van 19 tot 22°C. De vertakkingsgraad van de kiembuizen was positief gecorreleerd met de kiembuislengte.Kieming en appressoriumvorming verliepen beter op de bladschijven dan op glasplaatjes. Bij de behandelingen met variabele temperaturen werden significante correlaties waargenomen tussen de temperatuursom en de kiembuislengte en ook tussen de temperatuursom en het kiem- en appressoriumpercentage.De resultaten tonen aan dat kieming en vooral appressoriumvorming bij lagere temperaturen kan plaatsvinden dan verondersteld in de literatuur. Ze suggereren dat penetratie vanH. vastatrix het snelst gerealiseerd kan worden in de natuur als, na bevochtiging van de sporen, een temperatuurdaling plaatsvindt van aanvankeljk ca. 23 naar ca. 17°C binnen een paar uur. Deze conditie komt veelvuldig voor in de namiddag en het begin van de nacht in vele gebieden waar Arabica-koffie wordt verbouwd.     

一个编码推定的钙依赖磷酸酶催化亚基的番茄全长cDNA序列的克隆及其表达分析

 钙依赖磷酸酶(Calcineurin)作为Ca²⁺信号传感器中继因子对受Ca²⁺调节的生物学过程起重要调控作用。本研究克隆了一个番茄全长cDNA序列,该序列与钙依赖磷酸酶催化亚基(Calcineurin A,CNA)序列同源。推定的蛋白产物由315个氨基酸组成,明显短于非植物来源的该类蛋白。该蛋白含有钙依赖磷酸酶类金属磷酸酶中保守的催化作用活性位点基序,以及两个潜在的钙调蛋白结合位点,故暂时命名为LeCAL1(Lycopersicon esculentum calcineurin A like 1)。对CNA序列的分析结果表明,植物与非植物CNA差异明显,在系统进化树中形成距离很远的两大分支。植物CNA中,LeCAL1与拟南芥CNA同源性较高,而与水稻CNA较低。对LeCAL1在番茄中的表达分析结果显示,Cf/Avr介导的过敏性反应的产生诱导LeCAL1基因的表达,表明钙依赖磷酸酶可能在番茄抗叶霉病中起调节作用。     

水稻中与白叶枯病菌hrf1基因同源序列克隆及功能分析

利用PCR技术从粳稻R109中克隆到与hrf1同源的序列,命名为hpfr1,与hrf1基因的同源性为92%。推导的氨基酸同源性为74.8%。hpfr1基因连接到含T7启动子的高表达载体pET30a（＋）上构建重组质粒pHOSJ4,并转化宿主菌BL21（DE3）产生表达菌株BL21（DE3）/pHOSJ4。hpfr1与hrf1基因在起始密码子ATG至225个碱基处完全相同。将ATG至225碱基的序列克隆,并构建表达载体pHOSJ4-225,表达产物注射烟草能引起过敏反应。BL21（DE3）/pHOSJ4和BL21（DE3）/pHOSJ4-225的蛋白抽提物诱导烟草抗烟草花叶病毒病均达极显著水平,浓度为60μl/ml时防效最显著,分别为96.35%、95.4%,与harpinXoo防效（95.9%）相近。     

菜豆苗期抗锈性鉴定方法及种质资源的抗性筛选

多种环境因素影响病原菌的致病性和寄主的抗病性,从而对抗病性鉴定和筛选的准确性产生影响。苗期人工接种温度超过28℃,保湿时间少于6h,接种时连续光照等都影响菜豆锈病菌的萌发和侵入。接种浓度过低(<10~3个孢子/ml)或过高(>10~5个孢子/ml)、接种的苗龄过大(超过叶片展平期)及接种的不同部位(叶正、背面)也都对菜豆的抗性表现产生严重影响。菜豆锈病菌虽为专性寄生菌,但其保存方式简单(夏孢子保存于4℃冰箱内),且可较长时间保存(至少9个月)。作者根据这些影响因素提出了菜豆苗期抗锈性鉴定方法,并对我国260份菜豆种质资源进行了抗锈性筛选。结果表明,我国菜豆种质资源的抗锈性类型丰富,这一结果为菜豆的生产和抗病育种研究提供了可利用的品种或材料。     

水稻白叶枯病菌harpin基因的克隆与表达

 用PCR方法从水稻黄单胞菌白叶枯致病变种(Xanthomonas oryzae pv.oryzae,Xoo) JxoIII菌株中克隆到编码诱导植物过敏性反应激发子的基因hrfA。同源性比较发现其推测产物与水稻白叶枯病菌菲律宾小种Pxo86的Hpa1有97.2%的序列一致性,比Hpa1少了一个-GGG-GG-氨基酸残基序列重复。基因经NdeI和BamHI双酶切后连到含T7启动子的高表达载体pET 30a (+)上构建重组质粒pHRF4,并转化宿主菌BL21(DE3)产生表达菌株BLHR4。经过IPTG诱导之后,BLHR4产生一分子量为15.6 kD的蛋白质。研究表明,该蛋白在性质与功能上类似于其它已发现的harpins,即能够在烟草上引起典型的过敏性反应,富含甘氨酸、热稳定以及对蛋白酶敏感。因此,我们把该蛋白定名为harpin_Xoo。harpin_Xoo还具有诱导植物抗病性的功能。     

四川省小麦品种抗条锈现状及对策措施

近年来小麦条锈病在四川连年大发生。究其原因,最主要是大面积生产小麦品种抗锈性的丧失。分析本省小麦品种抗锈性现状,是新审定品种区试抗性水平有较大提高,但新品种一经大面积推广后抗锈性衰退现象十分严重。目前本省已采取多种有效措施,包括提高品种审定时的抗锈性指标,实行老品种"退役"制,加大品种更换的力度和合理品种布局,以及建立科学的抗锈性评价体系等。但要进一步提高本省小麦品种的整体抗锈性水平以控制小麦条锈病的发生和为害,还需要加强品种抗源及其抗性遗传背景的研究,加强学科间协作和对外交流,加大良种补贴面等。