拟康宁木霉Tk1的分离鉴定、拮抗作用及其生物学特性

为了分离利用高效植物病害生物防治菌株，从广东药用植物金丝皇菊根部分离获得一株木霉菌株Tk1。经过形态观察、rDNA-ITS和EF-1α序列分析明确其为拟康宁木霉Trichoderma koningiopsis。用平板对峙培养法测定，该菌株对金丝皇菊枯萎病菌（尖孢镰刀菌Fusarium oxysporum）和柑橘炭疽病菌（胶胞炭疽菌Colletotrichum gloeosporioides）的抑制率分别为68.5%和77.1%，且在后期能完全覆盖或部分覆盖病原真菌。其无菌发酵液对金丝皇菊枯萎病菌的抑制率为45.55%。该菌株营养生长和产孢的最适碳源分别为乳糖和葡萄糖，最适氮源是酵母膏；在pH 4~11的培养基上均可生长、产孢，最适pH为7；12 h交替光照条件下菌丝的生长速度最快，全光照条件下产孢数量最大；该菌株的致死温度为73℃处理10 min。     

花绒寄甲人工繁育及应用研究

为解决花绒寄甲人工繁育技术的关键问题，进一步提高产量，开展了低温保存对花绒寄甲卵活力的影响、替代寄主选择、成虫人工饲料配方、室内生活史等研究，并在后期初步开展了花绒寄甲对桃红颈天牛的野外防效试验。结果表明，4℃条件下保存0～60 d对花绒寄甲卵孵化率的影响存在显著差异，不经低温处理的卵孵化率最高，可达70.87%，低温保存60 d后，孵化率降为65.81%；大麦虫可作为花绒寄甲幼虫的替代寄主，最佳接虫比为1:5，此时化蛹率达70.40%，羽化率达94.03%；黄粉虫蛹、大蜡螟幼虫无法繁育出花绒寄甲成虫；黄粉虫幼虫干粉、大麦虫幼虫干粉、无菌水、脯氨酸、羟氨酸、丝氨酸、胱氨酸、葡萄糖、亚油酸混合配制的饲料使花绒寄甲成虫存活率达96.70%，单雌产卵量达161.10粒，产卵期持续43 d，是成虫的最佳人工饲料；经室内繁育的花绒寄甲一年发生2代，成虫寿命2～3年，卵期平均8.64 d、幼虫期平均6.63 d、蛹期平均36.88 d。花绒寄甲可用于防治桃红颈天牛，野外最佳释放比为8:1～ 10:1（天敌:天牛），虫口减退率达72.70%。     

柑橘砂皮病菌生防菌的筛选及其发酵条件优化

以柑橘砂皮病菌为靶标菌株,采用平板对峙法从土壤中分离到的227株细菌中获得4株对柑橘砂皮病菌有明显抑制活性的菌株,其中Y2菌株发酵液稀释5倍和10倍后,对柑橘砂皮病菌菌丝生长的抑制率分别为69.85%、61.76%。生理生化测定与16S r RNA基因序列分析结合,确定Y2菌株为短小芽孢杆菌(Bacillus pumilus)。Y2菌株培养的最佳氮源和最佳碳源分别为蛋白胨和可溶性淀粉,最佳发酵温度28℃,转速180 r/min。Y2菌株产生的抑菌活性物质经80℃高温处理和强酸、强碱处理,活性明显下降。     

具有除草活性的生防菌株GD-9发酵条件优化及菌剂制备

为了明确具有除草活性的生防菌株GD-9发酵过程中各因子的配比和最优条件,采用单因素试验对菌株最适碳源、氮源、载体、固态发酵基质进行了筛选,应用正交试验设计研究了碳源、氮源、初始含水量、接种量、初始pH、培养时间、培养温度7种因素对菌株活菌数的影响。试验结果表明菌株GD-9最佳固态发酵条件为:氮源NaNO_3 58.4 mg/g,碳源葡萄糖48.4 mg/g,培养基质最适初始含水量为258 mg/g,最适接种量为0.26 mL/g,初始pH 7.6,最佳培养时间147.8 h,最佳培养温度30.7℃,最适宜的载体为黏土,分散剂为聚乙烯醇,稳定剂为膨润土,润湿剂为糊精。通过发酵试验结果制备生防菌株GD-9的固体菌剂,该研究为菌剂的商品化生产奠定基础。     

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.     

棉花苗期根腐类病害生防细菌的筛选与鉴定

【目的】从新疆棉田中筛选对棉花苗期根腐类病害具有较强拮抗作用的生防细菌,为棉花苗期根腐类病害生物防治奠定基础。【方法】采用平板对峙法,测定实验室保存的多株生防细菌的抑菌能力,通过田间试验,验证其防治棉花苗期根腐类病害的效果。对生防细菌的16S rDNA基因序列与gyrA基因序列分别测序,通过BLAST在线比对,利用MEGA 7.0构建系统发育树,确定该生防菌株的分类地位。【结果】生防细菌TWt34、HWt34对棉花苗期根腐类病害(立枯病和红腐病)防治效果分别为76.74%和78.43%;在平板对峙对立枯丝核菌(Rhizoctonia solani) 和拟轮枝镰孢霉(Fusarium verticilliodes)的抑菌宽度分别为8.29和6.60 mm,具有较强拮抗作用,对尖孢镰刀菌(Fusarium oxysporum)和大丽轮枝菌(Verticillium dahliae)的抑菌宽度分别为7.70 和13.15 mm,也具有明显抑制作用。16S rDNA与gyrA基因序列鉴定表明生防菌菌株TWt34、HWt34同属于莫海威芽孢杆菌(Bacillus mojavensis)。【结论】筛选出的2株莫海威芽孢杆菌TWt34、HWt34对棉花4种主要病害的病原菌具有较强的抑制作用,能够有效防治2种棉花苗期根腐类病害(立枯病和红腐病)。     

芽孢杆菌微生态制剂对京红1号蛋鸡产蛋性能和蛋品质的影响

试验旨在研究芽孢杆菌微生态制剂对京红1号蛋鸡产蛋性能和蛋品质的影响。选择健康、体况良好的404日龄京红1号蛋鸡960只,随机分为4组,每组6个重复,每个重复40只。对照组饲喂基础日粮,试验Ⅰ组饲喂基础日粮+益倍佳(凝结芽孢杆菌),试验Ⅱ组饲喂基础日粮+赐尔健(纳豆芽孢杆菌),试验Ⅲ组饲喂添加地衣芽孢杆菌+枯草芽孢杆菌(1∶1)混合制剂的日粮,试验组微生态制剂的添加量均为300 mg/kg。试验期25 d,其中预饲期5 d。结果显示,与对照组相比,试验组蛋鸡产蛋率均显著提高(P<0.05),破、软蛋率均显著降低(P<0.05),试验Ⅰ、Ⅱ组料蛋比显著降低(P<0.05)。与对照组相比,Ⅱ组鸡蛋平均蛋重显著提高了2.25%(P<0.05);试验组Ⅰ、Ⅱ的蛋黄相对重、蛋壳相对重、哈氏单位均无显著影响(P>0.05);试验Ⅲ组蛋壳颜色为1~3级的鸡蛋比例明显增多,差异显著(P<0.05)。综上所述,添加微生态制剂赐尔健(纳豆芽孢杆菌)可显著提高京红1号产蛋率和平均蛋重,显著降低料蛋比和破、软蛋率;添加枯草芽孢杆菌+地衣芽孢杆菌(1∶1)混合制剂可使蛋壳颜色加深的鸡蛋比例显著增多。     

烟草青枯病生防菌的筛选及其田间防效评价

为筛选出对烟草青枯病有效防治的生防菌,通过稀释涂布法从烟株根际土壤分离到一株具有明显拮抗作用的菌株69-1,经16S rDNA鉴定该菌株为壮观链霉菌(Streptomyces spectabilis),并用该菌株进行烟草青枯病田间防治试验。结果表明,与对照相比,经菌株69-1处理烟株茎围、有效叶片数、最大叶宽、最大叶长、上中下部烟叶单叶干重、最大叶面积、产量均显著增加,且对烟草青枯病的相对防效达60.42%,同时对不同处理组的初烤烟叶进行化学成分的检测分析,发现菌株69-1处理相对于对照组烟叶品质更佳。该菌株菌剂处理不仅可以提高烟株对烟草青枯病的防效,而且还提高烟叶品质和产量,可为烟草青枯病生物防治提供一定参考。     

Diversity,plant growth-promoting traits,and biocontrol potential of fungal endophytes of Sorghum bicolor

The diversity of fungal endophytes in Sorghum bicolor was investigated in samples collected from 10 different geographical regions of Karnataka state, India. A total of 360 endophytes were isolated from leaf, stem, and root tissues and were assigned to 26 fungal species based on morphology and molecular characterization using ITS sequences. All the endophytes belonged to the phylum Ascomycota. The diversity (Shannon H, 2.57; Simpson_1-D, 0.92) and species richness (Margalef's, 4.68; Menhinick, 3.61) were found to be higher for the endophytes isolated from leaf tissues. The species evenness of the endophytic assemblage was strongly influenced by tissue type, followed by geographical location. The biocontrol potential of isolated endophytes was evaluated against economically destructive sorghum grain mould pathogens such as Fusarium thapsinum, Epicoccum sorghinum, Alternaria alternata, and Curvularia lunata using the dual culture method. Biocontrol potential was exhibited by 26 endophytic isolates, of which Trichoderma asperellum recorded broad-spectrum activity against target pathogens, followed by E. nigrum and A. longipes. Most (82%) endophytes showed plant growth-promoting traits. Biosynthesis of indole-3-acetic acid (IAA) was observed in 84% of isolates, and phosphate solubilization, siderophore production, and cellulase activity was observed in 69%, 23%, and 27% of isolates, respectively. Seeds treated with T. asperellum exhibited a significantly higher seed vigour index (2096), germination percentage (94%), and yield under greenhouse and field conditions. The results were substantiated by the confocal microscopy analysis, which clearly demonstrated the colonization of treated endophyte in root tissues. The present study reveals an ecofriendly approach to explore T. asperellum in sorghum disease management.