寡糖对向日葵锈菌侵染的影响

为了明确寡糖对向日葵锈菌侵染的影响,在向日葵苗期用寡糖处理并接菌后对锈菌夏孢子的萌发率和侵入率以及气孔开闭进行了观察,并用甲苯胺蓝染色法对入侵位点酚类化合物的积累进行了检测。试验结果表明,与对照相比寡糖处理后锈菌夏孢子的萌发率和侵入率均显著降低,而气孔关闭率显著升高。同时,在接种后12 h时,大量的侵入位点产生了酚类化合物的积累,说明寡糖处理可以有效抑制向日葵锈菌的侵染。     

向日葵不定根的研究──Ⅲ向日葵不定根的超微结构

利用透射电镜，对向日葵真根侧根及３种类型的不定根进行了超微结构的观察。结果表明，向日葵不定根与其真根侧根在导管类型上是一致的，均为网纹导管；菌根现象普遍存在；不定根维管组织细胞出现一定程度的细胞器优势。     

向日葵品种对锈病抗性的组织学和超微结构研究

为了进一步了解向日葵与向日葵锈病病菌的互作关系及品种的抗性机制,采用电子显微镜技术对向日葵品种对锈病抗性的组织学和超微结构进行研究.在电镜下观察发现,向日葵锈病菌侵染向日葵后,在感病品种和抗病品种上发育的组织学和超微结构特征有明显差异.病菌在感病品种细胞间隙分布有大量的胞间菌丝,寄主细胞发生质壁分离;叶绿体变形,叶绿体的片层结构排列零乱,直至叶绿体解体.在抗病品种上表现为菌丝生长受抑,没有观察到吸器的产生,寄主细胞的早期坏死与锈菌的发育受阻密切相关.品种的抗病性可能与饥饿阻止真菌生长有关.     

向日葵柄锈菌效应子P2的定位及功能研究

由向日葵柄锈菌（Puccinia helianthi Schw.）引起的向日葵锈病严重威胁着向日葵的安全生产。效应子是一类病原菌侵染过程中分泌的蛋白,促进病原菌侵染或触发寄主免疫。为明确向日葵锈菌效应子P2在锈菌侵染过程中发挥的作用,对该基因序列进行了生物信息学分析;以向日葵锈菌夏孢子cDNA为模板,克隆了P2基因的编码序列,利用qRT-PCR技术分析了该基因在侵染过程中的表达特性;采用农杆菌瞬时表达系统在烟草上验证了P2的毒性功能,并在烟草上瞬时表达对其进行了亚细胞定位。研究结果表明,P2编码58个氨基酸,N端含24氨基酸的信号肽,无核定位信号及跨膜结构域,并在向日葵锈菌侵染早期上调表达。在本氏烟中瞬时表达P2可以抑制BAX诱导的细胞坏死,亚细胞定位结果表明P2定位在细胞膜及细胞核。     

向日葵锈菌胞外金属蛋白酶的特性分析

为进一步了解锈菌侵染向日葵的致病机制,对向日葵锈菌(Puccinia helianthi)分泌的一种新型的金属蛋白酶(metalloproteinase,Mep)水解活性和生物学特性进行了表征。使用偶氮酪蛋白法测定蛋白酶的活性,在波长280 nm下测吸光度。结果表明,该蛋白酶对偶氮酪蛋白具有水解活性,纯酶最适温度和pH分别是50℃和7.0,在pH 5.0～9.0和温度50℃以下保持稳定。蛋白酶受到抑制剂乙二胺四乙酸二钠(EDTA)、乙二醇双(2-氨基乙基醚)四乙酸(EGTA)、1,10-菲咯啉的显著抑制;Ca²⁺、Zn²⁺、Mg²⁺可以显著提高该蛋白酶的活性,表明该酶为金属蛋白酶。另外,利用qRT-PCR试验测定了基因Mep在锈菌侵染向日葵的过程中的表达情况,结果显示,在接菌48 h时,基因表达量达到高峰,后逐渐降低,表明该蛋白酶参与了锈菌对向日葵的早期侵染过程。     

向日葵柄锈菌效应蛋白的预测及筛选

向日葵柄锈菌（Puccinia helianthi Schw.）引发向日葵锈病,对生产具有破坏性。为深入了解锈菌效应蛋白,研究病原与寄主的互作机理,在获得向日葵柄锈菌分泌蛋白组的基础上,利用生物信息学软件预测及分析向日葵柄锈菌效应蛋白,采用qRT-PCR检测其中7个候选效应蛋白基因在接种叶片中的相对表达量。结果表明：在供试的900个锈菌的分泌蛋白中预测到候选效应蛋白497个。基因特征分析显示,开放阅读框长度在200～399 bp之间的最多;信号肽长度集中在16～27个氨基酸残基（占92.96%）;信号肽氨基酸组成中亮氨酸出现频率最高,其次为丙氨酸、异亮氨酸;信号肽识别位点以SpⅠ型为主。本研究获得了7个向日葵柄锈菌的效应蛋白,qRT-PCR检测表明柄锈菌接种向日葵叶片后,与纯孢子相比,这7个效应蛋白编码基因均上调表达,说明其参与了锈菌侵染向日葵的过程。     

Susceptibility of US sweet corn hybrids to Puccinia sorghi Schw. in Hawaii

Ninety-eight sweet and supersweet corn hybrids, composites and inbreds were scored in two trials for resistance to common rust, Puccinia sorghi Schw., under epiphytotics at the Waimanalo Research Station of the University of Hawaii. The average rust rating on a 1–9 scale was very high (6·32), with scores ranging from 1·90 to 8·92. Only four hybrids and composites exhibited high resistance (rating ⩽2·5), of which the top two had been bred in Hawaii. An additional 18 hybrids exhibited moderate resistance (2·6–5·0). It is concluded that economic yield loss to rust of commercial sweet corn in the US could become much greater if breeding programmes are not strengthened.     

Yield and quality losses caused by common rust (Puccinia sorghi Schw.) in sweet corn (Zea mays) hybrids

Yield and quality of fresh ears were measured in field plots of selected hybrids of sweet corn (Zea mays L.) that were affected by rust (Puccinia sorghi Schw.) or were nearly rust-free (mancozeb-sprayed). In 1978, 28 hybrids were tested. Losses in total yield ranged from zero in the more resistant entries to nearly 50% in more susceptible entries. In 1979, three cultivars were planted. Yield losses of these cultivars were similar in ranking order to those in 1978, although the loss was greater in late-planted plots because of the greater final severity of the disease. Losses in total yield in late-planted sweet corn were 18%, 26% and 49% for cv. Sugarloaf (most resistant), cv. Jubilee (intermediate) and cv. Style Pak (most susceptible), respectively.     

寡糖诱导向日葵抗锈病的信号转导途径

为了解寡糖诱导向日葵抗锈病过程中的信号转导途径,利用寡糖、水杨酸（SA）、茉莉酸（JA）预先喷雾处理向日葵叶片,1d后接种锈菌,通过对植株体内脂氧合酶（LOX）活性及SA含量的测定,并通过RT-PCR对SA、JA信号途径的标记基因PR5和PDF1.2等表达水平进行测定,初步确定诱导抗性表达的信号转导途径。结果表明：经寡糖诱导后,向日葵叶片内LOX活性降低,SA含量上升,且诱导水杨酸途径标记基因PR5表达量增加。结果说明寡糖诱导向日葵抵抗锈病主要与SA介导的信号途径有关。     

Effect of crop management on epidemics of phomopsis stem canker (Diaporthe helianthi) for susceptible and tolerant sunflower cultivars

Phomopsis stem canker (Diaporthe helianthi) is a worldwide fungal disease which is responsible for high yield losses in sunflower crop in the main regions of production. Field data on the influence of crop management on the incidence and severity of stem canker were reported by Debaeke et al. (2003) but a more thorough study was required to analyse step-by-step the spread of the fungus within the canopy from spore deposition on leaves to stem injury.In a 2-year study (2000–2001), the effects of crop management (plant density, N fertilization, and irrigation) and genotypic tolerance (susceptible vs tolerant cultivars) on the epidemics of Diaporthe helianthi were monitored under conditions of reinforced inoculum. The incidence and severity of leaf and stem symptoms were closely related to canopy development (leaf area index) and microclimatic conditions (relative humidity) resulting from different crop management options. Increasing plant density resulted in a greater proportion of girdling stem lesions, detrimental to yield, because of earlier infection under dense canopies. The number of girdling lesions per plant was maximal with high N fertilization but more leaves were infected without N fertilization.The field data were used to evaluate satisfactorily the epidemiological Asphodel model regarding the main periods of leaf infection. The relative humidity within the canopy (observed), the number of leaf infection events (simulated by Asphodel) and the final proportion of stem lesions (observed) were positively related which clearly demonstrated the key role of crop management in the development of stem canker in sunflower.     

向日葵抗锈病基因同源序列的克隆与分析

向日葵锈病严重影响向日葵的产量。为了找到抗锈病相关基因,根据已知NBS-LRR型抗病基因保守结构域P-loop和GLPL设计简并引物,以接菌的抗病向日葵品种CM29叶片的cDNA为模板进行扩增。通过克隆转化得到10个具有连续开放阅读框的RGA,经系统进化分析将其分为TIR-NBS-LRR和non-TIR-NBS-LRR两种类型。对其氨基酸序列进行多重序列比对结果显示所获得的RGA具有典型的NBS-LRR型抗病基因保守结构域即P-loop、kinase-2a、kinase-3a和GLPL结构。运用BLASTX分析这些结果,表明其RGA与已知的抗病基因相应保守区域的同源性为18.1%～51.1%,说明它们可能与抗病功能基因具有密切联系。