Genetic variation among asexual progeny of Phytophthora infestans detected with RAPD and AFLP markers

Genotypic variation among 32 single-zoospore isolates (SZI) of Phytophthora infestans , derived asexually from two hyphal-tip parental isolates (PI-105 and PI-1) of the US-8 genotype, was assessed with 80 random amplified polymorphic DNA (RAPD) primers and 18 amplified fragment length polymorphic DNA (AFLP) primer pairs. In previous investigations, the SZIs from parental isolate PI-105 showed high levels of virulence variability and were differentiated into 14 races, whereas the SZIs from PI-1 showed identical virulence to the parent. The purpose of this investigation was to determine if phenotypic variation observed among SZIs of P. infestans could be detected at the DNA level in these isolates. Polymorphism was detected with 51 RAPD primers and with all 18 AFLP primer pairs in PI-105 SZIs. In SZIs from PI-1, polymorphism was also detected with 25 RAPD primers and 17 AFLP primer pairs. Cluster analysis using the unweighted pair-group method with arithmetic averages (UPGMA) separated the SZIs from parent PI-105 into six virulence groups, 11 RAPD groups and three AFLP groups. Cluster analysis of PI-1 SZIs, which all belong to the same virulence group, differentiated them into four RAPD groups and six AFLP groups. No close correlation among RAPD, AFLP and virulence groups could be established within the two progenies of SZIs. Results of this study suggest that there is a considerable level of inherent genetic variability among SZIs derived asexually from the same parental isolate. The possible mechanisms and implications of this genetic variation are discussed.     

Leaf, floret and seed infection of wheat by Pyrenophora semeniperda

Infection processes of Pyrenophora semeniperda on seedling and adult wheat leaves and wheat ears were investigated. Almost 100% germination of conidia occurred on seedling leaves, compared with 20–30% on adult leaves. Appressoria formed over the anticlinal epidermal cell walls and haloes always accompanied infection. Sometimes papillae formed within the leaves as a resistance mechanism. Infection hyphae ramified through the intercellular spaces of the mesophyll resulting in cellular disruption. The infection processes on floral tissues were similar to those observed on leaves; however, no infection occurred on anther, stigmatic or stylar tissues. Infection of ovarian tissue occurred both with and without appressoria formation. Hyphae grew mainly in the epidermal layers and appeared unable to breach the integumental layer as no growth was observed in endosperm or embryo tissues. The optimum dew period temperature for conidial germination was 23·6°C, compared with 19·9°C for lesion development, 20·4°C for the production of infection structures on seedling leaves and 23·7°C for floret infection. Leaf disease development occurred in a logistic manner in response to dew period, with maximum infection observed after 21 h compared with > 48 h in seeds. An initial dark phase during the dew period was necessary for infection and temperature after the dew period had an effect, with significantly more numerous and larger lesions being formed at 15°C compared with 30°C. Seedling leaves were found to be more susceptible than older leaves, under both field and controlled environment conditions. Infection of wheat seeds following inoculation of ears, or after harvest burial of inoculated disease-free seeds, was demonstrated. In the latter, 3-week-old seedlings were slightly stunted, whereas older plants were unaffected. The apparent unimportance of this plant pathogen as a cause of leaf disease in relation to its poor adaptation to dew periods and dew period temperature is discussed, along with the importance of its seed borne characteristics.     

A new approach to evaluate relative resistance and tolerance of tomato cultivars to begomoviruses causing the tomato yellow leaf curl disease in Spain

A simple procedure to evaluate relative resistance and tolerance of tomato cultivars to the begomoviruses causing tomato yellow leaf curl (TYLC) disease in Spain was developed. To estimate the resistance and tolerance levels of a cultivar, several formulae were developed based on the ratio of infected plants, virus titre (estimated by tissue–print hybridization) and symptom intensity. The formulae were applied to five commercial tomato cultivars (Amoretto, Birloque, Royesta, Tovigreen and Ulises) naturally infected by TYLC viruses. The analyses showed that Ulises, Birloque and Tovigreen exhibited a moderate resistance, and Ulises was also highly tolerant. There was a positive correlation between symptom intensity and virus titre in infected plants, suggesting that the hybridization technique could also be used as an early estimator of tolerance. Finally, molecular hybridization and nucleotide sequence analyses of the begomovirus intergenic region showed that the local TYLC virus population consisted of a single species, Tomato yellow leaf curl virus (TYLCV, formerly TYLCV-Israel), with low genetic variation (nucleotide identity between isolates higher than 97%).     

拮抗细菌菌株Bacillus spp.间的相互作用及其对水稻纹枯病的抑制

测定了6个Bacillus spp.和1个Pseudomonas sp.P-13(G-对照菌株)在PPM培养基上的相互作用。在总共42个组合中，有27个组合是可亲和性的。在以B5423为单剂之一的5个组合中，亲和性的组合有4个。在温室条件下通过分批播种，在水稻分蘖盛期喷以B5423-R(B5423的标记菌株，是利福平抗性的突变体)为单剂之一的4个亲和性组合的菌悬浮液，以检测该组合中B5423-R在水稻茎基部的种群数量。结果发现，与单剂B5423-R相比，在应用1～7天内，组合B5423-R B45中，B5423-R的群体数量显著性降低；组合B5423-R B95、B5423-R B77中，B5423-R的群体数量与单剂无显著差异；组合B5423-R B4313中，B5423-R的群体数量在应用1～4天内与单剂无显著性差异，但在应用第7天显著地高于单剂。离体叶片纹枯病痛斑面积的抑制试验表明，与各自的单剂相比，组合B95 B5423及B95 Er77显著地抑制了纹枯病病斑面积；而组合B45 B80、B45 B5423和B45 B95纹枯病病斑面积显著地高于各自的单剂；其余10个组合与各自的单剂无显著差异。     

哈茨木霉几丁质酶诱导及其对水稻纹枯病菌的拮抗作用

 在实验室条件下分别以几丁质和水稻纹枯病菌(Rhizoctonia solani)细胞壁作唯一碳源诱导哈茨木霉(Trichoderma harzianum)菌株NF9、TC3和P1产生几丁质酶,用硫酸铵沉淀法制备几丁质酶粗提液。上述木霉菌株内切几丁质酶活性(对胶体几丁质浑浊度的减少率)分别为79.8%、74.4%和76.0%,均显著高于非诱导的阳性对照。培养第5 d几丁质诱导的木霉菌株NF9和TC3内切几丁质酶活性显著高于由水稻纹枯病菌细胞壁诱导的酶活性。体外测定表明,通过诱导的木霉菌株TC3、NF9和P1几丁质酶粗提液对水稻纹枯病病菌的拮抗圈直径可达38、21和23 mm,与非诱导的阳性对照比较有显著性差异。对木霉几丁质酶拮抗作用的特点及生防应用进行了讨论。     

引起糖甜菜细菌性叶斑病的萎蔫短小杆菌新致病变种

 1995年在内蒙古临河市新发现了糖甜菜细菌性叶斑病,从病斑所分离的10个细菌菌株经柯赫氏法则验证,均确系该病的病原菌。采用形态观察、表型特征和生理生化特性测定、数值分析、血清学反应、细胞化学成分分析、DNA G+C mol%和DNA-DNA同源性测定进行了鉴定,并与植物病原棒形细菌15个标准菌株进行了比较。该病原菌为革兰氏阳性细菌,不规则短杆状,有一根鞭毛、亚极生或侧生,结合其生理生化特性、细胞化学成分和DNA G+C mol%和DNA-DNA同源性测定结果,认为应属于短小杆菌属(Curtobacterium)的萎蔫短小杆菌(Cur. flaccumfaciens),数值分析也支持这一结论。此外,据血清学反应结果及其对短小杆菌属的其它植物寄主的致病情况,认为该病原菌应是萎蔫短小杆菌种下的一个新的致病变种,定名为Curtobacterium flaccumfaciens pv. beticola pv. nov. Chen et al.,2000(萎蔫短小杆菌糖甜菜致病变种)。     

梨火疫细菌实时荧光PCR和诱捕PCR-ELISA检测方法的建立

根据梨火疫细菌中独特而保守存在的质粒pEA29，设计了1对引物和3条探针，建立了实时荧光PCR检测方法和诱捕PCR-ELISA检测方法。实时荧光PCR采用带荧光标记的核酸杂交探针，边扩增边检测，步骤简单，不需PCR后处理，可避免假阳性和交叉污染；诱捕PCR-ELISA检测方法只需简单处理的样品就能检测，减少了核酸不纯出现的漏检，由于增加了核酸杂交探针，可不需凝胶电泳EB染色检测，不会出现假阳性问题。     

种植方式与种植密度对大力士高粱的影响

本试验用裂区设计,研究了大力士高粱在16种不同种植密度、3种种植方式下,产草量、生产速度、茎叶比、茎粗的变化情况。结果表明,在撒播密度为18 kg/hm2、条播为22.5 kg/hm2、穴播为28.5 kg/hm2时达到最高产量;生长速度在第53-55 d时最高,达5.7 cm/d;茎叶比随着产量的增加逐渐减少;茎粗随着种植密度的增大而有减小的趋势。     

草莓摘叶处理对果实芳香物质的影响

 对草莓(Fragaria×ananassa Duch．)‘哈达’品种坐果后植株进行摘叶处理，对成熟果实的糖类和芳香物质含量进行了测定分析，结果表明：不摘叶、摘1/3叶和摘2/3叶3个处理果实GC/MS分析分别检测出43、33和37种芳香物质成分。随摘叶程度加重，芳香物质成分中酯类的相对含量呈下降趋势，而醛类的相对含量呈上升的趋势；2，5-二甲基-4-甲氧基-3(2H)-呋喃酮的相对含量明显降低；果糖和总糖含量显著降低。     

中国18 省市番茄晚疫病菌生理小种的鉴定

 利用5 个含有不同单显性抗番茄晚疫病基因的番茄材料Ts19、Ts33、W1Va700、LA1033 和L3708为鉴别寄主, 对我国18 省、市、自治区的番茄晚疫病菌201 个纯化分离物进行了生理小种鉴定。结果共鉴定出8 个小种, 即生理小种T0、T1、T1 ,2 、T1 ,2 ,3 、T1 ,2 ,3 ,4 、T1 ,4 、T1 ,2 ,4 和T3。其中, 小种T1 和T1 , 2是主流小种, 地理分布最广和发生频率最高, 分别在13 和11 个省市出现, 占样本总数的28.8 %和28.4 %;其次是小种T0 和T1 ,2 ,3 , 分别在8 和7 个省市出现, 分别占1819 %和819 %; 再其次是小1 ,2 ,3 ,4 , 在4个省市有发生, 占710 %; 小种T1 ,2 ,4 、T1 ,4 和T3 分布均少, 仅在1～2 个省市发生, 分别占3.0 %、2.5 %和2.5 %。这是我国的首次报道, 为番茄晚疫病的抗源材料筛选和抗病育种提供了病理学依据。