水稻白叶枯病菌转录调控因子基因fleQxoo和σ54因子基因rpoNxoo的分子鉴定

 为了阐明水稻白叶枯病菌(Xanthomonas oryzae pv.oryzae,简称Xoo)鞭毛生物合成及运动性的调控机理,本研究首先通过基因克隆、序列分析、缺失突变及表型测定,对转录调控因子fleQxoo和编码σ⁵⁴因子的基因rpoNxoo进行了分子鉴定。通过基因特异性扩增,成功地从Xoo菌株PXO99^A中克隆了fleQxoo和rpoNxoo。其基因序列与其它黄单胞病原菌中的同源序列高度保守。FleQxoo是NtrC家族激活蛋白成员之一,具有与σ⁵⁴作用的结构域和DNA结合保守结构域(HTH)。用标记交换法构建了△fleQxoo和△rpoNxoo基因缺失突变体。与PXO99^A相比,△fleQxoo和△rpoNxoo鞭毛产生能力丧失,运动性减弱,基因互补可以使之恢复;但其胞外纤维素酶和木聚糖酶活性以及对烟草叶片组织的致敏性无明显改变。因此,FleQxoo和σ⁵⁴主要参与了鞭毛生物合成及其运动性的调控。     

转录激活因子FleQxoo原核表达及其与鞭毛基体基因fliExoo启动子的结合

 为了阐明转录激活因子FleQxoo对水稻白叶枯病菌(Xanthomonas oryzae pv. oryzae,Xoo)鞭毛基体基因fliExoo的转录调控机理,本研究用PCR方法从Xoo野生型菌株PXO99^A中克隆了fleQxoo基因,构建了原核表达载体pET21-fleQxoo,在大肠杆菌BL21(DE3) pLysS中进行了诱导表达,通过Ni-NTA亲和层析纯化得到FleQxoo蛋白;EMSA检测到FleQ-xoo与鞭毛基体基因启动子fliExoo-p片段的特异结合作用。这些结果为阐明FleQxoo直接调控鞭毛基体基因fliExoo的转录提供了分子证据。     

寄主诱导下的玉米弯孢叶斑病菌致病性分化相关蛋白的动态变化研究

 Reinoculation and two-dimensional electrophoresis were performed to analyze the pathogenicity differentiation of Curvularia lunata.Resistant host inbred lines Shen135,Mo17,and 78599-1 were reinoculated(six generations) with low virulent isolate WS18.Results showed that the disease index had no significant change for the first 2 generations of inoculation.At the third generation,the incidence of disease was increased and the number of differential expressed proteins of mycelia were more than that in the first 2 generations.More than 100 differential expressed proteins were found in the mycelia of fifth generation when compared with the original one.In the experiment,10 differentially expressed proteins were identified by MALDI-TOF-MS analysis.Three proteins were related directly to the differentiation of virulence,2 were related to allergen,4 were related to the metabolism of carbon or signaling pathway and 1 was unkonwn to Curvularia lunata.     

安徽省烟草黑胫病菌的交配型及其地理分布研究

 Total 69 isolates of Phytophthora nicotianae var. nicotianae were isolated and identified from tobacco black shank samples collected from different areas of Anhui province, and mating types of the oomycete were investigated by the method of partnership culture in vitro on L-tryptophan medium with β-sitosterol. The results showed that A2 mating type was the predominant in isolates of P. nicotianae var. nicotianae, next was A₀ mating type, A₁ and other mating types were not observed. It was suggested that sexual reproduction of P. nicotianae var. nicotianae might not take place frequently, and Anhui province might not be the original center of P. nicotianae var. nicotianae. The results indicated further that mating type geographical distribution of P. nicotianae var. nicotianae in Anhui province was in line with that in China. In addition, the significance of the test results and the possible reason for A₁ mating type being absent were discussed.     

壳寡糖诱导烟草防御酶系活性变化及PR-1a基因表达研究

 The activity change of defensive enzymes and PR-1a gene expression of tobacco (Nicotiana tabacum) seedling induced by chito-oligosaccharides were studied. The results showed that high level systemic acquired resistance (SAR) was expressed in tobacco plants treated with chito-oligosaccharides solution at the concentration of 50 μg/mL. PAL activity increased greatly with 2 peaks, the activity of SOD decreased initially followed by an increase with higher increment, and the activity of POD peaked early followed by a gentle fall in chito-oligosaccharide treated plants. The PR-1a gene was strongly expressed in tobacco due to systemic acquired resistance induced by chito-oligosaccharides. At 168 h after inoculation the expression quantity (co-pies/2 μL) of PR-1a gene was increased to 2 469.6 in treated tobacco leaf, reached 392.6% than that at 0 h after inoculation, it was increased 3.05 times of that in untreated control.     

辣椒褐斑病菌分生孢子产生条件初探

 The sporulation conditions of Cercospora capsici were tested with physical and chemical methods. Conventional and particular media, ultraviolet radiation and different treatments on mycelia suspension were applied and studied. It was found that C. capsici could produce conidia in the media of corn leaf powder agar and pepper leaf powder agar. The numbers of conidia produced in the two kinds of botanical media were 24 000 and 35 000 spores/mL, respectively. Compared with those isolated from the leaves, the conidia produced in the botanical media were more slender and pointed in morphology.     

一个二粒小麦LRR类受体蛋白激酶基因的克隆和鉴定

 Near the HMW-glutenin gene of wheat (Triticum aestivum), there is a locus (temporarily named TaXa) encoding LRR-receptor-like protein kinase, which is homologous to disease resistance protein Xa21 of rice (Oryza sativa). Through RT-PCR approach, a cDNA clone of ZS2002 was isolated from the orthologous locus of TaXa in Triticum turgidum. ZS2002 was 3 081 bp long and encoding a peptide composed of 1 026 amino acid. The protein included N-terminal conserved sequence, LRR domains, a transmembrane region and a serine/threonine protein kinase domain. ZS2002 was expressed in root, stem, leaf and spike. The transcribing in seedling leaves was significantly enhanced by Blumeria graminis f.sp. tritici. TaXa gene might play a role in powdery mildew resistance reaction in Triticum.     

花生斑驳病毒青岛分离物cp基因序列克隆与分析

 Peanut mottle virus(PeMoV) was detected via RT-PCR from two peanut samples(QD5 and QD6) with mottle symptom collected from Qingdao, Shandong Province.The 3'-terminal 892 bp fragments of their genome were cloned and sequenced.The cp genes of QD5 and QD6 were 837 bp in length and encoded 278 amino acids(aa), with DAA at aa sites regulating aphid transmission.QD5 and QD6 shared nucleotide identities of 95.3%-99.4% and aa identities of 93.5%-99.6% in cp genes with other PeMoV isolates available in the GenBank.The phylogenetic results showed that PeMoV were clustered to three groups, America, Asia and Australia, which were consistent with their geographical origins.This is the first molecular evidence on the incidence of PeMoV in China.     

稻瘟病菌激发子诱导非寄主植物的抗病效果

 Hyphal cell wall crude elicitor(CWE)of rice blast pathogen could induce hypersensitive response in tobacco and induce other nonhost plants to be resistant to other fungal pathogens.When corn was treated with CWE,they inhibited the infection of Exserohilum turcicum and Curvularia lunata,and when plants of capsicum and cucumber were treated with CWE,they inhibited the infection of Colletotrichum gloeospori-oides.CWE solution showed no bioactivity on spore germination and hyphal growth of the experiment fungi in vitro.Nonhost resistance induced by CWE to other fungal pathogens was not complete resistance.The induced resistant effect(IRE) increased as CWE concentration increased,however,IRE had somehow satura-ted concentration of CWE.Induced nonhost resistance by incompatible pathogen was quantitative to other compatible pathogens.The induced resistance was best at 2 or 3 d after CWE treatment,and then decreased.IRE was about 20 percent in 10 d after CWE treatment.     

来自安徽不同寄主PVY分离物的血清学鉴定及其cp基因分子进化分析

 Tobacco and potato samples showing symptoms of PVY were collected from different regions in Anhui Province, and the ELISA results of partial samples were positive. The total RNA was extracted from the positive samples by TRIZOL methods. Specific primer pair was designed to amplify cp gene of PVY by RT-PCR. Sequencing results indicated that the full length of cp gene of PVY from tobacco (PVY-CP-4) and pota-to (PVY-CP-7) is 801 nts, and each of them encodes 266 amino acids. A phylogenetic tree based on alignment of cp nucleotide sequences was constructed and the sequence comparing of cp gene was conducted. The results showed that PVY-CP-4 could be grouped into one branch with PVY^O and PVY^N:O and shared the highest sequence similarity (99.4%) with PVY^O (EF026074). It was suggested that PVY-CP-4 derived from tobacco in Hefei might belong to PVY^O. PVY-CP-7 clustered together with PVYN and PVYNTN and formed another branch. Furthermore, PVY-CP-7 shared the highest sequence similarity (98.3%) with PVY^NTN(AJ890347), PVY^NTN (EF026075) and PVY^NTN(AJ585342). It was supposed that PVY-CP-7 derived from potato in Wuhe probably belonged to PVY^NTN.     

水稻纹枯病菌菌核上的真菌和细菌种类及其对水稻纹枯病菌生长的影响

 A total of 136 fungal and 86 bacterial isolates were isolated from the sclerotia of Rhizoctonia solani,collected from three different sources.These fungal isolates were identified as the following 12 genera:Alternaria,Aspergillus,Cladosporium,Coniothyrium,Curvularia,Gliocladium,Fusarium,Metarhizium,Peni-cillium,Phoma,Phytophthora and Trichoderma.Some isolates of Trichoderma and Gliocladium were found to inhibit the growth of R.solani strongly in vitro.Among the 86 bacterial isolates,20 of them could inhibit the growth of R.solani,whereas 5 isolates were found to promote the growth of mycelia and the formation of sclerotia of R.solani.These bacterial isolates with antagonistic actives to R.solani were identified as Bacillus and Pseudomonas.     

江西甘蔗花叶病病原的分子鉴定

 Sugarcane mosaic disease, caused by Sugarcane mosaic virus (SCMV), Sorghum mosaic virus (SrMV), Maize dwarf mosaic virus (MDMV) or Johnsongrass mosaic virus (JGMV) in Potyvirus, is one of the most important viral diseases of sugarcane. In the study, four primer pairs specific to SCMV, SrMV, MDMV and JGMV, respectively, were designed and used to detect 29 sugarcane leaf mosaic samples collected from 9 locations in Jiangxi province. The representative RT-PCR products were sequenced. The results showed that 22 samples were infected by SCMV, three by SrMV, and four were mix-infected by SCMV and SrMV. MDMV or JGMV were not identified in all samples. The result indicates that SCMV is the major pathogen of sugarcane mosaic disease in Jiangxi province, and SrMV is also a pathogen for the disease.     

Effects of short-term low temperatures on photosystem Ⅱ function of samara and leaf of Siberian maple (Acer ginnala) and subsequent recovery

Samara is the reproductive organ (seed) for many tree species in arid land in northwestern China. It is ecologically important in population development due to its dispersal function. However, information on its photosynthesis and effect of environmental stresses on its photosynthesis is still very limited. In the present study, responses of photosystem II (PSII) activity in samara and leaf of Siberian maple to short-term chilling/freezing and subsequent recovery potential were comparatively investigated by using polyphasic fluorescence test. The samara had more efficient photosynthesis (Fv/Fm and PIABS) and more efficient electron transport (φEo) but lower energy dissipation (DIo/RC) than leaf. Generally, the PSII performance and the electron transport for both samara and leaf were inhibited under low temperature stress, accompanied by an increase of energy dissipation in PSII reaction centers (RCs). PSII of both samara and leaf was not markedly affected by chilling and could acclimate to chilling stress. Short-term freezing could completely inhibit PSII activity in both samara and leaf, indicated by the drop of values of Fv/Fm, PIABS, φEo to zero. PSII functional parameters of short-term dark frozen samara could be largely recovered whereas those of frozen leaf could not be recovered. The higher tolerance of samara to short-term low temperature stress than leaf is of great ecological significance for seed development, population establishment of Siberian maple.     

Relationship between dew presence and Bassia dasyphylla plant growth

Dew has been recognized for its ecological significance and has also been identified as an additional source of water in arid zones.We used factorial control experiment,under dew presence in the field,to explore photosynthetic performance,water status and growth response of desert annual herbage.Bassia dasyphylla seedlings were grown in contrasting dew treatments (dew-absent and dew-present) and different watering regimes (normal and deficient).The effects of dew on the water status and photosynthetic performance of Bassia dasyphylla,grown in a desert area of the Hexi Corridor in Northwestern China,were evaluated.The results indicated the presence of dew significantly increased relative water content (RWC) of shoots and total biomass of plants in both water regimes,and enhanced the diurnal shoot water potential and stomatal conductance in the early morning,as well as photosynthetic rate,which reached its maximum only in the water-stressed regime.The presence of dew increased aboveground growth of plants and photosynthate accumulation in leaves,but decreased the root-to-shoot ratio in both water regimes.Dew may have an important role in improving plant water status and ameliorating the adverse effects of plants exposed to prolonged drought.     

玉米大斑病菌孢子萌发和附着胞形成的影响因素研究

 Factors of influence on conidium germination and appressorium formation of Setosphaeria turcica,such as light condition,conidial concentration,nutrient resources and pH value of conidial suspension were studied.There was no significant difference among light treatments.The optimal pH for conidium germination and appressorium formation was 5.0 to 7.0.The exogenous nutrient sources were not the indispensable factors for conidium germination,but 5% sugar solution were more favorable for appressorium formation than the others.Low conidial concentration in suspension (conidia ≤ 10⁴/mL) was propitious to conidium germination and appressorium formation,which were inhibited significantly in higher concentration.It was suggested that the phenomenon was due to the self-inhibitor,a kind of lipophilic substance,existing in the site of conidium germination.     

来源于沿叶脉褪绿症状桃的PLMVd分离物群体结构分析与序列测定

 Peach latent mosaic disease occurred in peach trees is distributed widely in China. The disease shows a wide range of symptoms in the fields and discoloration along leaf veins is easily found. To get insight into the correlation between the symptom and Peach latent mosaic viroid (PLMVd), leaves of a peach tree showing the symptom of discoloration along its veins were collected and subjected to PLMVd RNA extraction, and RT-PCR detection. The target amplified fragment was cloned, and some positive clones were then selected for analyzing population structure of the isolate by single-strand conformation polymorphism (SSCP) analysis. Three predominant clones of the isolate were sequenced and analyzed. This study would provide more information for understanding the correlation between the molecular characterization of PLMVd and the symptom of discoloration along leaf veins.     

外源茉莉酸和真菌激发子诱导白菜CaMBP10的表达

 Recently,CaMBP10 was identified as a plant lipid transfer protein(LTP).In the study,four leaves old cabbages were treated with exogenous jasmonic acid(JA),JA plus LTP,fungal elicitor,200 mmol/L NaCl and 20% PEG respectively for understanding the functions of CaMBP10 in vivo.The expression of CaMBP10 was determined by using semi-quantitative RT-PCR.The results showed that the expressions of CaMBP10 were dramatically up-regulated in different levels under indicated conditions.It demonstrated that CaMBP10 was involved in biotic and abiotic stress response.     

水稻白叶枯病菌鞭毛基体基因fliExoo缺失突变分析

 为了阐明水稻白叶枯病菌(Xanthomonas oryzae pv. oryzae,简称Xoo)鞭毛基体组分蛋白基因fliExoo的生物学功能,本研究用Gm^R抗性基因标记交换法成功构建了基因缺失突变体△fliExoo。与野生型菌株PXO99^A相比,△fliExoo鞭毛缺失,菌体易沉降,在0.3%半固体培养基上运动能力明显降低;尽管生长速率和胞外纤维素酶活性无明显变化,但胞外多糖产生和生物膜形成能力明显下降;对水稻品种日本晴的致病性和对非寄主烟草的致敏性反应明显减弱。基因互补可以使上述突变表型恢复。因此,鞭毛基因fliExoo突变不仅影响了细菌鞭毛依赖的运动性,而且对病原菌毒性相关的表型也产生了影响。     

分子标记辅助选择小麦抗白粉病兼抗赤霉病聚合体

 Sumai 3, a wheat variety resistant to Fusarium head blight(FHB), was crossed with Neimai 9, a commercial wheat cultivar with the resistance to powdery mildew.The SCAR(sequence characterized amplified region) markers of powdery mildew resistance gene Pm21 and four SSR(simple sequence repeats)markers flanking the major FHB resistance QTL(Qfhs.ndsu-3BS) in Sumai 3 were used to detect the resistance loci by marker assisted selection(MAS) in the plants of the F₂ population.Identification of resistance to both powdery mildew and FHB in field showed that 12 plants resistant to both diseases were obtained.In addition, the agronomic traits of these plants were better than those of Sumai 3, and are perhaps the excellent parental materials for wheat breeding.