Trichoderma Biocontrol of Colletotrichum acutatum and Botrytis cinerea and Survival in Strawberry

Trichoderma isolates are known for their ability to control plant pathogens. It has been shown that various isolates of Trichoderma, including T. harzianum isolate T-39 from the commercial biological control product TRICHODEX, were effective in controlling anthracnose (Colletotrichum acutatum) and grey mould (Botrytis cinerea) in strawberry, under controlled and greenhouse conditions. Three selected Trichoderma strains, namely T-39, T-161 and T-166, were evaluated in large-scale experiments using different timing application and dosage rates for reduction of strawberry anthracnose and grey mould. All possible combinations of single, double or triple mixtures of Trichoderma strains, applied at 0.4% and 0.8% concentrations, and at 7 or 10 day intervals, resulted in reduction of anthracnose severity; the higher concentration (0.8%) was superior in control whether used with single isolates or as a result of combined application of two isolates, each at 0.4%. Only a few treatments resulted in significant control of grey mould. Isolates T-39 applied at 0.4% at 2 day intervals, T-166 at 0.4%, or T-161 combined with T-39 at 0.4% were as effective as the chemical fungicide fenhexamide. The survival dynamics of populations of the Trichoderma isolates (T-39, T-105, T-161 and T-166) applied separately was determined by dilution plating and isolates in the mixtures calculated according to the polymerase chain reaction (PCR) using repeat motif primers. The biocontrol isolates were identified to the respective species T. harzianum (T-39), T. hamatum (T-105), T. atroviride (T-161) and T. longibrachiatum (T-166), according to internal transcribed spacer sequence analysis.     

梨叶片中苹果褪绿叶斑病毒的引物原位标记检测

 选用带苹果褪绿叶斑病毒的香梨叶片,制备成适合进行原位RT-PCR的石蜡切片。设计特异引物,利用引物原位标记技术对切片组织中的病毒进行了检测研究,结果表明,PRINS-FITC染色法和PRINS-Rhodamine染色法均能获得良好的检测效果,有病毒部位分别显示绿色荧光和红色荧光,而且荧光出现的位置与原位RT-PCR检测的结果一致。由此证明引物原位标记技术可用于果树病毒的原位检测。     

梨树苹果茎痘病毒PRINS检测中引物的设计及退火温度的测定

[目的]利用5条引物目的片段进行扩增,完成PRINS检测中引物及其退火温度的测定.[方法]采用确认带有ASPV的库尔勒香梨叶片,以叶片为试验材料,并采用经RT - PCR检测技术健康香梨的叶片作为阴性对照.引物采用NCBI提供的BLAST工具中的ASPV病毒的全基因组序列的外壳蛋白的保守区域进行设计.并根据PRINS特有的性质及ASPV病毒有片断相互重叠现象来设计相应的引物.[结果]这5条引物之间没有太多的干扰,且都在目的区域出现了比较清晰的条带.[结论]证明所设计引物可以在进一步的PRINS检测ASPV病毒时直接使用,为PRINS在梨树病毒检测方面发挥更大的作用奠定了基础.     

蔬菜保护地木霉菌rDNA-ITS序列和UP-PCR遗传多样性分析

采用传统形态学分类和ITS序列比对的方法,研究蔬菜保护地土壤中木霉菌种群分布和遗传多样性。木霉菌分离培养结果显示,共获得397株木霉菌,鉴定出11个种,分别为:长枝木霉Trichoderma longibrachiatum、深绿木霉T.atroviride、哈茨木霉T.harzianum、粘绿木霉T.viren、微孢木霉T.minutisporum、拟康木霉T.pseudokoningii、黄绿木霉T.aureoviride、非钩木霉T.inhamatum、棘孢木霉T.asperellum、长孢木霉T.longipile和螺旋木霉T.helicum。经ITS序列建立系统发育树后,将木霉菌分为5个组。用5条通用引物经UP-PCR扩增后,扩增出46条谱带,其中多态性条带43条,占总条带数的93.5%。遗传多样性分析表明,当相似系数为0.80时,可将24个菌株划分为9个组。UP-PCR与ITS序列相比,更能体现木霉菌种间和种内的亲缘关系及遗传差异性,可以作为木霉菌分类的辅助方法。     

催芽型包衣种对漂浮育苗中烟苗生长的影响

通过苗期试验，以烤烟品种K326，云烟85为材料，比较了催芽型烤烟包衣种和常规包衣种对漂浮育苗中出苗时间、出苗率以及根系、叶片干鲜重、根系活力等的影响．结果表明：催芽包衣种能显著提高漂浮育苗烟苗的出菌率，特别对提高烤烟幼苗出苗的速率和整齐度，缩短出苗时间等方面有较好的效果，催芽包衣种平均比常规包衣种提前3d左右出苗．催芽包衣种可以增加烟苗的株高，使茎秆伸长，促进根系的生长发育．无论是地上部还是地下部的物质积累量，催芽包衣种的烟苗都明显高于常规包衣种的；催芽包衣种对漂浮育苗烟苗的根系活力有明显的增加作用。     

鸡球虫病疫苗研发新思考

虽然球虫激发宿主产生的保护性免疫应答机制已有很多相关研究,但是保护记忆性免疫应答机制这一关键点常常被忽视。高效保护记忆性免疫应答是球虫感染宿主和球虫病疫苗设计的基础。本研究对该领域50年的相关研究进行系统性回顾和分析,提出开发高效球虫疫苗的设想和评估候选球虫病疫苗产生高效免疫保护的新策略,该设想和新策略基于产生高效保护性记忆免疫应答的三大关键点,即产生免疫水平的阈值、效应部位以及时间。结果表明:1)球虫病活卵囊疫苗与球虫重组蛋白疫苗、DNA疫苗、以病毒或细菌为载体的球虫病活载体疫苗免疫鸡群激发宿主产生的免疫应答存在本质区别,活卵囊疫苗免疫过的鸡对球虫的感染处于“Primed”免疫状态(即免疫系统可快速高效地识别球虫百余个抗原,抵抗球虫再次感染的能力更高。),而非活卵囊疫苗对球虫的感染仍处于“Naive”状态(即免疫系统仅能识别特定的1个或几个球虫抗原,抵抗球虫再次感染的能力较弱。)。这也是目前研发的基于一个或几个抗原的球虫“死”疫苗效果不佳的根本原因。2)高效的保护性记忆免疫应答的主要机制是多种效应机制协同发挥作用,如肠道微环境中的组织常驻记忆细胞(Tissue resident memory cells,T_RM)、循环的效应记忆细胞(Effector memory T cells,T_EM)、中心记忆性T细胞(Central memory T cell,T_CM),尤其是T_RM的量和质起关键作用。在疫苗设计中可以将抗原靶向可激发宿主产生针对鸡球虫高效保护性记忆免疫应答的重要部位—肠道。3)抑制球虫早期裂殖生殖是记忆性免疫应答抵抗球虫感染最重要的特征,为评估高效球虫疫苗提供了新思路。综上,对鸡球虫感染过程中产生高效免疫保护的机制有清晰的认识和理解,是实现基于免疫学理论的球虫病科学防控的基础。     

修剪对紫薇主干生长的影响

连续3年对速生紫薇、玫瑰红紫薇、普通紫薇树体进行不同修剪技术应用,发现修剪可显著增加主干的生长。尤其是冬剪加夏剪的处理,速生紫薇、玫瑰红紫薇、普通紫薇两年平均地径分别比相应的对照品种提高了34.4%、46.9%、58%,不同品种对修剪反应的敏感程度不同。修剪还可以改良主干干形,同时通过集中处理修剪枯枝落叶,对抑制病虫害也有一定的作用。     

CAPS DNA Marker Labeling Using Primed in Situ Hybridization Technique for Chromosome of Strawberry (Fragaria × ananassa Duch.)

The cleavage amplified polymorphic sequence (CAPS) markers were developed to identify strawberry cultivars from unknown samples. These markers seemed to have a chromosome specificity. However, no reports confirmed it. The present study was conducted to verify the location of a CAPS marker on the chromosomes using a primed in situ (PRINS) technique. A CAPS marker was hybridized with the ‘Sachinoka’ chromosomes using the PRINS technique. The samples were observed under a fluorescence microscope. When the chromosomes were hybridized with the single marker, fluorescent signals were found on two chromosomes. From the results, the CAPS marker was confirmed to have a chromosome specificity by chromosome observation.     

随机引物PCR技术及其在动物医学中应用的研究进展

随机引物PCR技术(AP-PCR)是一种建立在PCR基础上的新的DNA多态性检测技术,其特点是利用随机寡核苷酸引物扩增基因组DNA片段,在无需预知研究对象的情况下对其进行基因特征的分析,具有简便、高效、实用等优点,现已广泛应用于分子生物学的多个领域。文章主要对AP-PCR技术的原理、优缺点、研究近况及其在动物医学中的应用等作一综述。