内生菌-植物联合修复污染土壤研究进展

环境污染对生态系统及人类健康造成严重威胁。近年来,许多学者研究发现内生菌联合植物修复体系对修复自然环境中的重金属和有机物具有巨大的潜力。本文主要从内生菌联合植物进行污染修复的机理和应用两方面入手,介绍了内生菌接种宿主植物根、茎、叶部的几种方法及定殖的检测方法,总结了污染土壤中内生菌在植物组织内的定殖动态,以及内生菌-植物联合体系强化有机物污染和重金属污染修复效果的应用,并从内生菌调节生长因子、生物固氮和溶磷、在宿主植物体内的共代谢有机物、产生特异性酶降解有机物、提升植物对重金属抗性和降低重金属毒性几个方面解释了内生菌和植物的联合修复机理。最后指出,虽然已经有许多对于内生菌联合植物修复体系的研究,但是目前两者相互作用机理尚未完全清楚,对于多种内生菌组合体系以及处理水体和大气污染的研究并不完善,这些都将成为今后的研究重点。     

黄瓜疫霉菌拮抗内生细菌的筛选与鉴定

【目的】从黄瓜叶片中筛选抗疫霉菌效果好的内生细菌并进行鉴定,为进一步研究黄瓜疫病的理 论探索及防治应用提供实验材料。【方法】以黄瓜叶片为实验材料筛选内生细菌,根据待检菌株的镜检特征、 革兰氏染色反应、菌落排列方式和菌落形态特征等进行初步筛选。采用平板对峙法和发酵液拮抗法实验对目标 内生细菌进行拮抗作用测定。通过 16S rDNA 序列测定及同源性分析,并构建系统发育树确定目标菌株种属。 【结果】初步筛选出 32 株内生细菌将其编号为 CL1~CL32,其中革兰氏阴性杆菌 10 株、革兰氏阳性杆菌 13 株、革兰氏阳性球菌 9 株,利用平板对峙法发现 8 株对掘氏疫霉菌有较强抑制作用的拮抗菌株,菌丝抑制率为 10.3%~54.3%,抑菌半径为 1~7 mm,对其中抑菌效果较好的 3 株菌株（CL7、CL9、CL15）进行发酵液拮抗实验, 结果显示其抑菌率分别为 54.4%、72.2%、63.3%。【结论】对掘氏疫霉菌具有较强抑制作用的 3 株内生细菌（CL7, CL9、CL15）分别属于泛菌属 Pantoea sp.、假单胞菌属 Pseudomonas sp. 和葡萄球菌属 Staphylococcus sp.,其中 抑制效果最明显的为 CL9,即假单胞菌属 Pseudomonas sp.。     

植物内生菌对柑橘溃疡病的生物防治作用及应用前景

从植物内生菌的抗病机理及生物防治病害作用方面综述了利用植物内生菌防治柑橘溃疡病的机理与优势,并对其生物防治应用前景进行了展望。     

植物内生菌在害虫治理中的应用

对植物内生菌在害虫治理中的应用及其控制害虫的机理等方面的研究概况进行综述，并讨论了植物内生菌直接利用存在的主要问题和在害虫综合治理中的应用前景。     

生防菌R-4的鉴定及其对玉米南方锈病的防效

对分离自玉米植株的内生细菌R-4进行鉴定,测试其对玉米南方锈病(SCR)的防病增产效果。用盆栽方法测定菌株R-4对玉米植株的促生效果,根据16S r DNA序列对菌株R-4进行鉴定,采用自然发病方式测试其对SCR的防病增产效果。结果表明,菌株R-4处理玉米的发芽率、播种后18 d株高、播种后28 d株高、植株地上部干重及根干重比对照分别增加17.19%、32.03%、41.80%、92.53%和37.03%。通过16S r DNA序列分析,将菌株R-4鉴定为蜡样芽孢杆菌(Bacillus cereus)。菌株R-4在先玉335、银糯1号、浚单20上对SCR的防效达39.2%~92.1%(平均为65.7%),经该菌处理后玉米增产2.58%~9.02%(平均为6.66%)。     

利用变性梯度凝胶电泳分析正红菇菌根围土壤真菌群落多样性

以正红菇（Russula griseocarnosa）菌根围土壤为研究对象,通过提取土壤基因组DNA,以通用引物扩增真菌18S rRNA基因V1+V2区,将PCR产物进行变性梯度凝胶电泳（Denaturing Gradient Gel Electrophoresis）,获得土壤微生物群落的DNA特征指纹图谱,并对图谱中的优势条带回收测序,通过Blast进行同源性比对并构建系统发育树,进而分析正红菇菌根围真菌群落组成及多样性。同源性比对结果表明,在回收测序的19条DGGE条带中,4条为非真菌真核生物序列,系统发育分析显示全部序列可以分为4类菌群,GroupⅠ主要为担子菌门（Basidiomycota）真菌,GroupⅡ主要为子囊菌门（Ascomycota）真菌,GroupⅢ为未知真菌,GroupⅣ主要为节肢动物门生物（Arthropoda）。     

Identification of sources of resistance to damping-off and early root/hypocotyl damage from Rhizoctonia solani in common bean (Phaseolus vulgaris L.)

Rhizoctonia solani causes economically important root and hypocotyl diseases in common bean throughout the world. Root health is a vital factor in plant development and root diseases would negatively influence water and nutrient uptake as well as cause direct stand reduction and root rot damage to the crop. An efficient common bean screening method to evaluate damping-off and early root/hypocotyl damage from R. solani was developed and used to identify dry bean lines with levels of resistance to this disease. Two sets of 163 and 111 lines previously evaluated for drought tolerance in Nebraska and Puerto Rico were evaluated for damping-off resistance and early root/hypocotyl damage under greenhouse conditions. Disease severity on plants was identified based on above-ground symptoms, seedling survival and root lesions using a rating scale of 1 (resistant) to 9 (susceptible). In the first set of lines representing commonly grown dry bean cultivars, germplasm and sources of damping-off resistance, the Rhizoctonia mean rating ranged from 1.7 to 3.9; Phaseolus vulgaris lines PI 310668 and PI 533249 had the highest damping-off resistance. In the second set of the best lines from a drought tolerance shuttle breeding program the Rhizoctonia mean rating was between 2.6 and 5.7. The availability of drought tolerant dry bean lines allowed the testing of the hypothesis that there was a correlation between selecting for drought tolerance and R. solani damping-off resistance. No correlation between mean disease rating and drought tolerance was found, but adapted dry bean lines such as NE14-08-176 released as SB-DT1, and NE14-08-225 were identified with moderate damping-off resistance and drought tolerance. Lines with both traits and other attributes will facilitate development of resistant bean cultivars to manage damping-off caused by R. solani.     

Occurrence of endophytes in grasses native to northern China

A survey of grass species associated with Neotyphodium and Epichloë fungal endophytes was made in the permanent grasslands of northern China. A total of 41 grass species, 56 sites and 172 local populations were examined to reveal endophytic infection of native grasses. Twenty‐five of the forty‐one species of grasses (proportionately 0·61) were infected by endophytes: twenty‐two species had a mean infection rate of 0·01–0·50 and only three species had a mean infection >0·50. Of the 172 local populations examined, however, proportionately 0·72 were not infected, 0·181 had an infection rate of 0·01–0·50 and 0·099 had an infection rate of 0·51–1·00. Two genera of grasses, Cleistogenes and Koeleria, as well as twenty plant species, were previously unknown endophytic hosts in China. High infection rates (0·86–1·00) of plants of Achnatherum sibiricum were detected at all eleven sites studied. Other grasses in which high infection rates were found were Agropyron mongolicum, Roegneria turczaninovii, Agropyron elongate, Poa palustris and Poaangustifolia. It was concluded that endophytes are found widely in natural grass populations and that endophyte–grass interactions may have effects on ecosystems and livestock in the grasslands of northern China.     

野生黄花美冠兰根部内生真菌多样性研究

以海南省白沙县野生黄花美冠兰（Eulophia flava）作为试验材料,采用培养基培养方法分离、纯化并鉴定真菌,研究不同生境下黄花美冠兰植株根部的内生真菌群落多样性及其受生境的影响程度。结果表明,从丛生于甘蔗林和荒山中的黄花美冠兰植株新鲜营养根段中共分离出52株内生真菌,鉴定为17个属,丝核菌属（Rhizoctonia,13．46％）和镰刀菌属（Fusarium,13．46％）为优势属。相同生境的黄花美冠兰内生真菌群落丰富度较为一致;而两种不同生境中的黄花美冠兰内生真菌群落丰富度则表现出较大的差异,生于荒山的黄花美冠兰内生真菌群落Shannon多样性指数远高于生于甘蔗林的,可认为黄花美冠兰内生真菌多样性受生境的影响较大。     

茶树根系耐铝内生真菌的筛选鉴定

茶树是典型的喜铝耐铝植物,其根系微生物在铝元素的吸收转化中起重要作用.从茶树根系分离鉴定出一株耐铝的内生真菌,进行了显微镜形态鉴定及18S rDNA序列分析,分析了该菌株对不同重金属的耐受程度,研究铝对该菌生长的影响,并利用电感耦合等离子原子发射光谱仪(ICP-AES)对该菌株耐受A13+进行了研究分析.结果 发现,显微镜形态观察及18S rDNA序列表明该菌为粘红酵母属粘红酵母(Rhodotorula glutinis),命名为A3.粘红酵母A3对不同重金属的耐受性为:Al3+＞ Mn2+＞ Cu2+,其中铝浓度可达250 mmol·L-1.当培养基中添加了150 mmol·L-1的Ap+,菌液pH值为2.0～-3.0,粘红酵母A3生长良好.ICP-AES结果显示该菌株铝离子含量与培养基中Al3+浓度呈正相关,说明该菌株在一定范围内对Al3+具有吸附作用;同时测定该菌株的含水量随培养基中铝含量增加而增大,揭示该菌株对铝毒害的规避与其通过含水量的增加平衡细胞液有关系.分离的耐铝真菌为茶树的耐铝机制提供新的思路,并为其定殖到其他作物规避铝胁迫提供微生物资源,以提高植物的抗性,增加产量.