贡柑黑腐病病原菌生物学特性研究

摘要：在贡柑黑腐病病原菌分离鉴定的基础上,对其病原菌的生物学特性进行研究。结果表明,该病原菌（Alternaria citri Ellis & Everhart）在PDA培养基上生长最佳。菌丝生长和分生孢子萌发的适宜温度分别为21~30 ℃和18~30 ℃,适宜pH值分别为5~12和4~7。供试的6种碳源中,葡萄糖和可溶性淀粉较利于菌丝生长,蔗糖较利于孢子萌发;供试的6种氮源中,牛肉膏较利于菌丝生长,酵母膏较利于孢子萌发。光照条件对病原菌产孢影响不大,完全光照与光暗交替12 h较利于菌丝生长和孢子萌发。分生孢子致死温度为51 ℃水浴10 min。     

贡柑黑腐病病原菌的分子鉴定

贡柑黑腐病是近年来在贡柑上发生的一种新型病害,该病害可导致嫩梢快速枯死,大量嫩叶脱落,严重影响树势和产量。本研究从病区采集发病枝叶,对贡柑黑腐病病原菌进行分离、纯化,并采用rDNA-ITS序列分析技术对其进行分子鉴定。结果表明,贡柑黑腐病病原菌为链格孢属（Alternaria Nees）真菌,rDNA-ITS序列与GenBank中芸薹链格孢菌(Alternaria brassicae (Berk.) Sacc.)和柑桔链格孢菌(Alternaria citri Ellis & Everhart)的ITS序列的同源性为99%。结合形态学特征,推断该病原菌为柑桔链格孢菌。     

不同pH条件下接种菌根真菌对柑橘砧木苗生长及光合效应的影响

本文以枳橙（Citrus sinensis (L.) Osbeck譸oncirus trifoliate (L.)）为试验材料,利用盆栽试验,研究不同pH条件下接种丛枝菌根(Arbuscular mycorrhiza)真菌摩西球囊霉(Glomus mosseae)对柑橘砧木枳橙苗的生长及植株光合效应的影响。试验选定3个pH范围（pH 5.5~6.5, 6.5~7.5, 7.5~8.5）和接种摩西球囊霉为试验因子,共设6个处理组合,以不同pH条件下未接种摩西球囊霉为对照。采用单株小区,随机区组排列,重复10次。结果表明,接种摩西球囊霉能有效地侵染枳橙植株,菌根侵染率65%以上。试验还发现,处理植株的菌根侵染率和株高、生物量干重均显著高于对照植株,同时,接种AM真菌显著增加了植株叶片叶绿素含量,并显著提高处理植株光合作用速率和蒸腾速率。研究结果表明,接种AM真菌增强了宿主植物对土壤酸碱度的适应性,在酸性和碱性土壤上均能显著提高柑橘的光合效应。     

10种杀细菌剂对火龙果细菌性茎腐病菌的毒力测定

采用抑菌圈法,对10种杀细菌剂在火龙果茎腐病病原上进行了毒力测定。结果表明：72%农用硫酸链霉素可溶粉剂、53.8%氢氧化铜水分散颗粒剂（杜邦TM）、45%代森铵水剂、47%春雷?王铜可湿性粉剂、50%福美双可湿性粉剂和3%中生菌素可湿性粉剂对火龙果茎腐病病原均有一定抑制效果。其中,45%代森铵水剂和50%福美双可湿性粉剂防治效果较好,抑制率分别达到78.88%和71.42%,其直线回归方程分别为y=2.380 2.477x,y=2.896 1.146x,EC50分别为0.4929,0.0240。72%农用硫酸链霉素可溶粉剂效果次之,抑制率达到66.67%,直线回归方程为y=1.020 1.040x,EC50值为2.5686。这3种药剂可作为火龙果茎腐病田间药效试验的首选药剂。     

江西永修凤凰山桃实腐病病原鉴定

桃实腐病是江西永修凤凰山桃园发生的一种新的重要病害,其主要危害挂果期的果实,严重影响桃产量和品质。病果主要表现为脐部变褐腐烂,并在病部表面密生黑色小点（分生孢子器）,病果易脱落。本文对病果进行了组织分离,对获得的病菌纯培养进行了回接,同时对病菌的形态及大小观察和测量,结果表明引起凤凰山桃实腐病的病原菌为核果果腐拟茎点菌（Phomopsis amygdalina Canonaco）。     

一株马铃薯干腐病生防菌的鉴定及其抑菌活性研究

镰孢菌引起的马铃薯干腐病是影响马铃薯贮存期间产量与品质的重要病害之一。寻找高效低毒的防治马铃薯干腐病的生防药剂对提高马铃薯的产量与质量具有重要意义。本研究以腐皮镰孢Fusarium solani为病原菌, 测定分离自马铃薯块茎的菌株2-1-9-CJK-2的抑菌活性。该菌株发酵液、无菌发酵液、菌悬液、胞内组分以及挥发性有机物对腐皮镰孢均具有抑菌活性, 抑制率分别为93.37%、61.92%、80.82%、44.01%和49.16%。结合形态学、生理生化特征和多基因系统发育树构建鉴定其为贝莱斯芽胞杆菌Bacillus velezensis。马铃薯切片试验和马铃薯块茎试验表明, 当菌株发酵液浓度为1×109 cfu/mL时, 对病原菌的抑制效果最好。除此之外, 菌株可以分泌蛋白酶、淀粉酶、纤维素酶, 具有溶磷能力和生物膜形成能力, 但不具有溶血性, 并且对供试的其他6种病原真菌有较强的抑菌活性。研究表明贝莱斯芽胞杆菌2-1-9-CJK-2抑菌谱广, 具有生物安全性, 对腐皮镰孢有较强的抑菌活性, 在植物病害防治中具有一定的应用潜力, 可为马铃薯干腐病生防制剂的开发奠定基础。     

辣木内生真菌的分离及其抗菌活性物质的初步研究

从辣木中分离得到1株内生真菌ly14,其发酵液对金黄色葡萄球菌、绿脓杆菌和掷孢酵母有很强的抗菌作用．该菌发酵液中的抗菌物质对温度、pH及紫外线照射有较强的稳定性;ly14代谢物的理化特性表明,抗菌物质为极性酸性化合物,其紫外可见吸收光谱在276nm处出现了一个较为明显的吸收峰．     

昆虫病原真菌在害虫防治中对天敌生物的影响研究进展

昆虫病原真菌是一类重要的微生物杀虫剂,广泛应用于农林害虫的生物防治。由于寄主范围广和主动侵染的特点,昆虫病原真菌在侵染、致死靶标害虫的同时,对包括天敌昆虫在内的非靶标节肢动物等会产生潜在的致病或亚致死风险。本文总结了昆虫病原真菌在害虫防治中的研究进展及对天敌生物的影响,并展望了昆虫病原真菌作为极具潜力的生物农药与天敌生物联合防治害虫的未来发展方向。     

Screening of antagonistic Trichoderma strains and their application for controlling stalk rot in maize

Maize is one of the major crops in China, but maize stalk rot occurs nationwide and has become one of the major challenges in maize production in China. In order to find an environment-friendly and feasible technology to control this disease, a Trichoderma-based biocontrol agent was selected. Forty-eight strains with various inhibition activities to Fusarium graminearum, and Fusarium verticillioides were tested. A group of Trichoderma strains(DLY31, SG3403, DLY1303 and GDFS1009) were found to provide an inhibition rate to pathogen growth in vitro of over 70%. These strains also prevented pathogen infection over 65% and promoted the maize seedling growth for the main root in vivo by over 50%. Due to its advantage in antifungal activity against pathogens and promotion activity to maize, Trichoderma asperellum GDSF1009 was selected as the most promising strain of the biocontrol agent in the Trichoderma spectrum. Pot experiments showed that the Trichoderma agent at 2–3 g/pot could achieve the best control of seedling stalk rot and promotion of maize seedling growth. In the field experiments, 8–10 g/hole was able to achieve over 65% control to stalk rot, and yield increased by 2–11%. In the case of natural morbidity, the control efficiency ranged from 27.23 to 48.84%, and the rate of yield increase reached 11.70%, with a dosage of Trichoderma granules at 75 kg ha~(-1). Based on these results, we concluded that the Trichoderma agent is a promising biocontrol approach to stalk rot in maize.     

A novel glycoside hydrolase 74 xyloglucanase CvGH74A is a virulence factor in Coniella vitis

Grape white rot is a destructive fungal disease occurring worldwide. Recently, Coniella vitis was identified as the predominant pathogen causing this disease in China. As the periderms of grape shoots are severely degraded by C. vitis, it was speculated that cell wall-degrading enzymes (CWDEs) might play a key role in the pathogenesis of this disease. Therefore, this study aimed to examine the hydrolytic activity of the CWDEs of C. vitis. The results showed that xylanase (Xy) and xyloglucanase (XEG) had high levels of hydrolytic activity both in vitro and in vivo. Furthermore, a high-virulence fungal strain exhibited higher levels of Xy and XEG activities compared with a low-virulence strain. The genome of the fungus was found to harbor two XEG-coding genes CvGH74A and CvGH74B, which belonged to the glycoside hydrolase (GH)74 family. The expression level of CvGH74A was found to be high during pathogen infection. CvGH74A gene deletion mutants were generated using the split-marker method. The deletion of CvGH74A decreased both the hydrolytic activities of XEG and Xy and also the ability of the fungus to infect the grape leaves. No differences in the hyphal growth, morphology of colonies, or conidiation were found between the ΔCvGH74A mutant strains and the wild-type strain. Together, these results suggested that CvGH74A acted as an important virulence factor, and its enzymatic activity might regulate the virulence of the pathogen. This study was novel in reporting that GH74 XEG acted as a virulence factor in C. vitis.