Diversity of Paenibacillus durus strains isolated from soil and different plant rhizospheres evaluated by ARDRA and gyrB-RFLP analysis

Strains belonging to Paenibacillus durus isolated from the rhizosphere of various grasses and from bulk soil were previously divided into five phenotypic groups (A1–A5) based on the fermentation pattern of six carbohydrates (A1: sorbitol (+), A2: dulcitol and tagatose (+), A3: starch and glycogen (+), A4: starch, glycogen and d-arabitol (+) and A5: negative for these carbohydrates). This study aimed to assess whether plant types select for specific P. durus phenotypic groups. For that purpose, polymerase chain reaction-restriction fragment length polymorphism analysis of part of genes encoding 16S rRNA (ARDRA) and DNA gyrase subunit B (gyrB-RFLP) were used to produce genetic fingerprints. ARDRA and gyrB-RFLP data were clustered together to generate a dendrogram and two main clusters were observed. Cluster I showed a predominance of strains isolated from wheat, maize and sugarcane rhizospheres. Strains isolated from maize were distributed among the five patterns of carbohydrate metabolism, while strains isolated from sugarcane showed to be predominantly able to metabolize starch and glycogen. Neither sorbitol- nor arabitol-metabolizing strains were found in cluster II, which consisted of strains isolated from soil and from all plant species used. Our results suggest that the plants influenced the diversity of P. durus in their rhizospheres.     

多粘类芽孢杆菌对木麻黄青枯病的抑菌防病作用

室内测定了商品化生物农药多粘类芽孢杆菌可湿性粉剂对木麻黄青枯病病原菌的毒力。采用打孔法结果表明,多粘类芽孢杆菌对木麻黄青枯病病原菌具有良好的拮抗作用,其抑菌圈的平均直径达到9.4 mm。采集4个品系的木麻黄小枝水培生根后进行温室盆栽,采用改进的伤根淋菌法进行接种,以只施多粘类芽孢杆菌可湿性粉剂(CK1)和只接种青枯菌(CK2)为对照,施用多粘类芽孢杆菌可湿性粉剂后接种青枯菌为处理进行实验。结果表明,在两个时间点,湛江-3的防效达到了60.8%和51.6%;湛江-2的防效达到了41.5%和40.3%;广东A14的防效达到了28.8%和23.3%,差异均显著(α=0.05);广东701-3的防效为16.3%和26.8%,差异不显著,出现此原因是广东701-3抗性强于其他3个品系。综上所述,在盆栽试验中,多粘类芽孢杆菌可湿性粉剂对木麻黄青枯病有显著的防治效果,品系本身越感病,防效越高。     

一种新型丁醇产生菌的鉴定及发酵条件初探

[目的]对一株新型丁醇产生菌进行鉴定,并研究其发酵条件。[方法]通过富集培养、分离筛选纯化微生物菌株,利用气相色谱仪检测分离菌株的丁醇产量,并利用形态学观察、生理生化分析及16S rDNA分子鉴定确定分离菌株的种类。[结果]试验得到一株高产丁醇菌HIT1,其发酵时适宜的葡萄糖浓度为25 g/L,适宜的发酵初始pH为6.5,且与酵母粉相比,其能更好地利用蛋白胨作为氮源;试验确定HIT1为多粘类芽孢杆菌(Paenibacillus polymyxa)。[结论]该研究在一定程度上拓宽了产丁醇菌种的资源库,为综合开发利用生物丁醇奠定了菌种多样性基础。     

多粘类芽孢杆菌CP7对荔枝霜疫霉菌的抗菌活性及其作用机制

多粘类芽孢杆菌Paenibacillus polymyxa CP7菌株分泌的抗菌物质对荔枝霜疫霉菌等真菌具有很强的抑杀作用。用光学显微镜、扫描电镜和透射电镜观察发现,荔枝霜疫霉菌菌丝体经CP7菌株的抗菌物质处理后, 菌丝发生不规则扭曲、萎蔫、干瘪,菌丝端部膨大等形态变化;经处理的菌体细胞组织结构受损,细胞膜结构模糊不清,看不到完整的外膜结构,细胞器消溶。处理后的荔枝霜疫霉菌孢子囊出现凹陷、干瘪、畸形;孢子囊外壁严重受损,原来厚实的外壁只剩一薄层,孢子囊内原有的微细结构消失,结构模糊不清,细胞器溶解,孢囊内物质大量外泄。抗菌物质还引起菌体细胞内磷的外泄,并对细胞呼吸速率和NADH氧化酶活力有抑制作用。     

Fusaricidins in Paenibacillus polymyxa A21 and their antagonistic activity against Botrytis cinerea on tomato

Four kinds of antifungal compounds from an extract of Paenibacillus polymyxa A21 with molecular masses of 883.56, 897.59, 947.55, and 961.58 Da were characterized as the members of fusaricidin-type of antibiotics according to LC-MS analysis. Fusaricidins isolated from culture filtrate displayed high antagonistic activity against several plant fungal pathogens, especially Botrytis cinerea, the causal agent of gray mold. The fusaricidins biosynthetic gene cluster (BGC) from A21 was cloned by PCR and comparative cluster analysis revealed that gene fusTE, the 3′ boundary of the fusaricidin BGC in strain PKB1, was not present in fusaricidin BGC of A21, indicating that fusTE is not necessary for fusaricidin synthesis. Fusaricidin extract from A21 significantly reduced gray mold disease incidence and severity on tomato. The mRNA levels for three patho-genesis-related proteins (PRs) revealed that treatment of tomato leaves with fusaricidin extract induced the expression of PR genes to different levels, suggesting that one reason for the reduction of gray mold infection by fusaricidin is induction of PR proteins, which lead to increased resistance to pathogens. This is the first report of the application of fusaricidins to control tomato gray mold and the comparative cluster analysis provides important molecular basis for research on fusaricidin biosynthesis.     

3％井冈霉素A·(2亿芽胞/毫升)多粘类芽胞杆菌水剂防治水稻纹枯病试验

以3％井冈霉素A·(2亿芽胞/毫升)多粘类芽胞杆菌水剂为供试药剂,通过田间试验研究不同剂量、不同施药时期对水稻纹枯病的防治效果.2年试验结果表明,当用药量在45～90 g.a.i/hm2 时,药后20 d,发病初期(即病株率在5％以下)预先防治的田间防效达80.38％～89.06％,而发病期(即病株率在10％以上)施药的田间防效仅达41.12％～53.26％;在发病初期预先防治效果要明显优于发病期再施药,从而确定该药剂防治水稻纹枯病时期应适当前移.     

多粘类芽孢杆菌对小麦赤霉病田间防治效果研究

以郑麦9023为试材,研究了微生物农药多粘类芽孢杆菌对小麦赤霉病的田间效果。结果表明:5亿CFU/g多粘类芽孢杆菌悬浮剂400~600mL/667m~2在小麦抽穗初期施第1次药,间隔5d施第2次药,对小麦赤霉病防治效果较好,防效在80%以上,增产效果达14.51%~17.27%,可以广泛用于小麦赤霉病的防治。     

多粘芽孢杆菌中2,3-丁二醇脱氢酶的克隆和表达

将来自于多粘芽孢杆菌(Paenibacillus polymyxa)中的2,3-丁二醇脱氢酶基因pla C在大肠杆菌(Escherichia coli)BL21中进行了诱导表达,并对其酶学性质进行研究。结果表明,该基因的表达产物主要是生成内消旋型2,3-丁二醇,其氧化2,3-丁二醇的最适温度是50℃,最适pH为10,而还原乙偶姻的最适温度为60℃,最适pH为6。     

Biological control agents (BCAs) of verticillium wilt: influence of application rates and delivery method on plant protection,triggering of host defence mechanisms and rhizosphere populations of BCAs

Verticillium dahliae causes severe yield reductions in a variety of important annual crops worldwide. Control of verticillium wilt has relied on soil fumigation; however, the use of the main soil fumigant, methyl bromide, has been banned in the European Union since 2010, creating a demand for novel crop protectants. As such, the use of biocontrol agents (BCAs) is an appealing management strategy. Prerequisites for the development of a successful BCA are an understanding of the modes of action of the antagonist, its ecological fitness and an efficient and economically feasible delivery system. Therefore, two BCAs (Paenibacillus alvei K165 or the nonpathogenic Fusarium oxysporum F2) and two release strategies (seed coating or amendment of the transplant soil plug) were assessed against verticillium wilt of aubergine (eggplant). Mixing the transplant soil plug with K165 or F2, at a rate of 10 and 20% (v/v), respectively, reduced verticillium wilt symptom development. Furthermore, a positive correlation was revealed between the release strategy and the BCA rhizosphere population. Correlation analysis also showed that disease severity was negatively correlated to the rhizosphere size of the BCA population. In addition, qPCR analysis showed that both BCAs induced the expression of the pathogenesis‐related (PR) proteins PR1 and PR4 in the stem of aubergines before and after inoculation with V. dahliae in a manner that suggests a link with the rhizosphere size of the BCA population.     

多粘类芽孢杆菌对油菜中硝酸盐含量的影响

蔬菜中硝酸盐含量超标危害人体健康,有效降低蔬菜中硝酸盐含量是提高蔬菜品质的主要目标之一。本研究考察了多粘类芽孢杆菌对降低油菜中硝酸盐含量的作用。结果表明,在种植油菜的基质中施加尿素后,油菜中硝酸盐含量显著提高,远超出国家标准限值（3000 mg/kg）;而施加尿素后再施加多粘类芽孢杆菌发酵液,油菜中硝酸盐的含量保持在较低的水平,与不施尿素的对照相当;将发酵液高温灭活后,减少油菜中硝酸盐积累的作用消失。施用100倍至10000倍稀释的多粘类芽孢杆菌发酵液（3×105~3×107 CFU/mL）,对油菜中硝酸盐的积累都有明显降低作用,浓度越高效果越明显。