摘要
2011年3月、6月、9月和12月在河北省安国市中药材基地分0～10、10～20和20～30cm3个土层采集金银花根围土样，应用PCR-DGGE与克隆测序技术相结合的方法研究AM真菌遗传多样性及其与土壤因子的关系。结果表明，金银花根围AM真菌最佳DGGE条件为80g·L-1的丙烯酰胺、PCR产物上样量25μL、变性剂范围40％～60％、电泳时间8h、电压130V、温度60℃。以DGGE共分离出15条AM真菌条带，从中选取10条清晰粗亮的优势条带进行测序，通过同源性比对发现，10个序列均与未培养球囊霉属菌种相似性高（≥95％），说明球囊霉属为优势菌群。系统发育分析表明，10条优势条带聚为5个系统发育种，系统发育种在季节和土层上差异显著，但4个季节均未出现特有种。RDA分析表明，影响AM真菌分布的主要土壤因子是全氮、有机碳和pH。其中，AM真菌遗传多样性与土壤全氮和有机碳呈显著负相关，与pH呈显著正相关。
AM fungal genetic diversity in the rhizosphere of Lonicera japonica and the influencing soil factors were investigated by the methods of PCR-DGGE combined with molecular cloning techniques. Soil samples were collected in the rhizosphere of L. japonica and divided a depth of 30 cm into three sections, i. e. 0-10, 10--20 and 20--30 cm from Anguo city of Hebei province in March, June, Sep- tember and December 2011, respectively. DGGE was performed on polyacrylamide gels with the dena- turing gradient from 40% to 60% and 80 g .L-1 Acrylamide, 25 μL PCR products were used for DGGE analysis. The electrophoresis was running at a fixed voltage of 130 V for 8 h at 60 ℃. The fin- gerprint of DGGE showed that a total of 15 species of AM fungi were discovered. 10 bands of domi- nant species from DGGE fingerprint atlas were sequenced; all of the sequences had the highest similar- ity （≥95%） to uncultured Glomus. The result indicated that Glomus was the dominant genera of AM fungal colony in the rhizosphere of L. japonica. DGGE band sequences and phylogenetic analysis re- vealed that all sequences can be divided into five groups, the dominant species had definite differences among the seasons and soil layers, none of endemic species to four seasons. RDA analysis revealed that the distribution of AM fungi were significantly and negatively correlated with soil total N and or- ganic C, and significantly and positively correlated with soil pH.
出处
《西北农业学报》
CAS
CSCD
北大核心
2014年第2期114-119,共6页
Acta Agriculturae Boreali-occidentalis Sinica
基金
河北省自然科学基金(C2010000273)
